Use and pharmaceutical composition of phenylisoxazolyl methylene-naphthalene-ether derivatives

ABSTRACT

Provided are a method of treating or preventing infection with hepatitis B virus in a human or animal, comprising administering to the human or animal in need thereof a therapeutically effective amount of a phenylisoxazolyl methylene-naphthalene-ether derivative having a structure of formula (I); use of a phenylisoxazolyl methylene-naphthalene-ether derivative having a structure of formula (I) in the preparation of a pharmaceutical composition for anti-hepatitis B vims and a pharmaceutical composition for anti-hepatitis B virus.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a National Stage of International PatentApplication No: PCT/CN2020/120370 filed on Oct. 12, 2020, whichapplication claims the benefit of priority to International PatentApplication No: PCT/CN2019/122595 filed on Dec. 3, 2019, which is hereinincorporated by reference in their entirety.

TECHNICAL FIELD

The present invention relates to the field of pharmaceuticals fortreating or preventing hepatitis B virus (HBV) infection. Specifically,the present invention relates to the use of phenylisoxazolylmethylene-naphthalene-ether derivatives for the treatment or preventionof HBV infection, pharmaceutical compositions comprisingphenylisoxazolyl methylene-naphthalene-ether derivatives and otheranti-HBV agents, and pharmaceutical use of phenylisoxazolylmethylene-naphthalene-ether derivatives.

BACKGROUND ART

Hepatitis B virus (HBV) is a hepatotropic, enveloped, partiallydouble-stranded DNA virus. It is most commonly spread from mother tochild at birth (perinatal transmission) and also can be transmitted byblood or body fluid.

HBV generates a covalently closed circular DNA (cccDNA), secrets HBVsurface antigen to suppress the immune system, and caused persistent(chronic) infection which is hard to eradicate. HBV infection is a majorpublic health threat in the world with over 257 million peoplechronically infected and caused over 887000 deaths every year. (Revill,P. A. et al, Lancet Gastroenterol. Hepatol. 2019, 4(7), 545-558). In theAsia-Pacific region, chronic hepatitis B virus (HBV) infection causedmore than half of the deaths due to liver cirrhosis and about half thecases of hepatocellular carcinoma in the region (Sarin, S. K. et alLancet Gastroenterol. Hepatol., 2020, 5(2):167-228). In a meta-analysisof 27 studies, it was reported that the pooled estimated prevalence ofHBV infection in the general population of China from 2013 to 2017 was6.89% (Wang, H. et al BMC Infect Dis 2019, 19(1), 811).

Currently, treatments for HBV infection are very limited. Approvedtreatments include nucleot(s)ide inhibitors, such as Tenofovirdisoproxil (Viread), Tenofovir alafenamide (Vemlidy), Entecavir(Baraclude), Telbivudine (Tyzeka or Sebivo), Lamivudine (Epivir-HBV,Zeffix, or Heptodin) and immunomodulators, such as pegylated Interferonalfa-2a (Pegasys). Thus, novel treatments are urgently needed. (Fanning,G C, Nat. Rev. Drug Discov, 2019, 18(11), 827-844).

Farnesoid X receptor (FXR) is a member of the nuclear receptor family,which includes steroid receptors, retinoid receptors, and thyroidhormone receptors (Radreau P, Porcherot M, Ramière C, et al. Reciprocalregulation of farnesoid X receptor α activity and hepatitis B virusreplication in differentiated HepaRG cells and primary humanhepatocytes. FASEB J. 2016; 30(9):3146-54). FXR is mainly expressed inthe liver, kidney, small intestine and adrenal glands. After HBV infectscells, HBV cccDNA enters the nucleus and begins to transcribe a varietyof HBV RNAs. The pregenomic RNA is reverse-transcribed to form HBV DNA,and the remaining RNAs are translated to form HBsAg, HBeAg, HBcAg, HBx,etc. These viral proteins and nucleic acids are assembled together inthe endoplasmic reticulum, and then virus particles are formed and arereleased out of cells. In the life cycle of HBV, the transcription ofHBV cccDNA can be regulated by nuclear receptors, such as HNF-4α andFXR. In vitro experiments have confirmed that FXR agonists inhibit theformation of stable transcription complexes between cccDNA and HBx byactivating FXR, thereby affecting the stability of cccDNA, inhibitingthe transcriptional activity of cccDNA, and achieving the purpose ofreducing viral DNA and HBsAg (Mouzannar K, Fusil F, Lacombe B, et al.Farnesoid X receptor-α is a proviral host factor for hepatitis B virusthat is inhibited by ligands in vitro and in vivo. FASEB J. 2019;33(2):2472-2483).

DISCLOSURE OF THE INVENTION

The inventors of the present invention found that the phenylisoxazolylmethylene-naphthalene-ether derivatives represented by formula (I), asan efficient small molecule FXR agonist, has anti-HBV activity. In vitroprimary human hepatocyte infection test results show that thephenylisoxazolyl methylene-naphthalene-ether derivatives represented byformula (I) can effectively inhibit HBV DNA, HBV RNA and HBsAg.

Thus, the present invention relates to the use of phenylisoxazolylmethylene-naphthalene-ether derivatives for the treatment or preventionof HBV infection, pharmaceutical compositions comprisingphenylisoxazolylmethylene-naphthalene-ether derivatives and otheranti-HBV agents, and pharmaceutical use of phenylisoxazolylmethylene-naphthalene-ether derivatives.

Definitions

For purposes of interpreting this specification, the followingdefinitions will apply and whenever appropriate, terms used in thesingular will also include the plural, and vice versa.

As used herein, the term “C1-6 alkyl” denotes an alkyl radical havingfrom 1 up to 6, particularly up to 4 carbon atoms, the radicals beingeither linear or branched with single or multiple branching, forexample, butyl, such as n-butyl, sec-butyl, isobutyl, tert-butyl;propyl, such as n-propyl or isopropyl; ethyl or methyl; moreparticularly, methyl, iso-propyl or tert-butyl.

As used herein, “C1-6 alkoxy” refers to “C1-6 alkyl-O—”, and isparticularly methoxy, ethoxy, isopropyloxy or tert-butoxy.

As used herein, the term “C3-6 cycloalkyl” refers to a cyclic alkylradical having 3 to 6 carbon atoms, for example cyclopropyl, cyclobutyl,cyclopentyl or cyclohexyl. The C3-6 cycloalkyl can be optionallysubstituted by C1-6 alkyl and/or halogen.

As used herein, the term “C4-7 alkylcycloalkyl” refers to a combinationof alkyl and a cycloalkyl group such that the total number of carbonatoms is 4 to 7. For example, C4 alkylcycloalkyl includesmethylenecyclopropyl.

As used herein, the term “5-10 membered aryl” refers to a 5-10 memberedmonocyclic- or bicyclic- or tricyclic-aromatic ring system. Typically,the aryl is a 5 or 6 membered ring system.

As used herein, the term “5-10 membered heteroaryl” refers to a 5-10membered monocyclic- or bicyclic- or tricyclic-aromatic ring systemhaving 1 to 4 heteroatoms. Typically, the heteroaryl is a 5 or 6membered ring system. Furthermore, the term “heteroaryl” as used hereinmay encompass monovalent or divalent heteroaryls.

As used herein, the term “halogen” or “halo” refers to one or more offluoro, chloro, bromo and iodo, and more particularly, fluoro or chloro.

As used herein, the term “C1-6 haloalkyl” refers to an alkyl radicalthat is substituted by one or more halo radicals, and is particularlyC1-6 fluoroalkyl or C1-6 chloroalkyl, such as trifluoromethyl and2,2,2-trifluoroethyl.

As used herein, the term “pharmaceutically acceptable salts” refers tosalts which are not biologically or otherwise undesirable.Pharmaceutically acceptable salts include both acid and base additionsalts.

As used herein, the term “pharmaceutically acceptable auxiliarymaterials” may include any or all solvents, dispersion media, coatings,surfactants, antioxidants, preservatives (e.g., antibacterial agents andantifungal agents), isotonic agents, absorption delaying agents, salts,drug stabilizers, binders, excipients, disintegration agents,lubricants, sweetening agents, flavoring agents, dyes, and the like andcombinations thereof, as would be known to those skilled in the art.Except insofar as any conventional carrier is incompatible with theactive ingredient, its use in the therapeutic or pharmaceuticalcompositions is contemplated.

As used herein, “hepatitis B virus” or “HBV” refers to a member of theHepadnaviridae family having a small double-stranded DNA genome ofapproximately 3,200 base pairs and a tropism for liver cells. “HBV”includes hepatitis B virus that infects any of a variety of mammalian(e.g., human, non-human primate, etc.) and avian (duck, etc.) hosts.“HBV” includes any known HBV genotype, e.g., serotype A, B, C, D, E, F,and G; any HBV serotype or HBV subtype; any HBV isolate; HBV variants,e.g., HBeAg-negative variants, drug-resistant HBV variants (e.g.,lamivudine-resistant variants; adefovir-resistant mutants;tenofovir-resistant mutants; entecavir-resistant mutants; etc.); and thelike.

As used herein, the term “therapeutically effective amount” refers to anamount of a compound or molecule of the present invention that, whenadministered to a subject, (i) treats or prevents the particulardisease, condition or disorder, (ii) attenuates, ameliorates oreliminates one or more symptoms of the particular disease, condition, ordisorder, or (iii) prevents or delays the onset of one or more symptomsof the particular disease, condition or disorder described herein. Thetherapeutically effective amount will vary depending on the compound,the disease state being treated, the severity of the disease treated,the age and relative health of the subject, the route and form ofadministration, the judgment of the attending medical or veterinarypractitioner, and other factors. The present invention relates to apharmaceutical composition comprising an HBsAg inhibitor and anucleos(t)ide analogue, in a pharmaceutically acceptable carrier.

Unless specified otherwise, the term “compound of formula (I)” includesphenylisoxazolyl methylene-naphthalene-ether derivatives of the formula(I), prodrugs thereof, salts of the compound and/or prodrugs, hydratesor solvates of the compound, as well as all stereoisomers (includingdiastereoisomers and enantiomers), tautomers isotopically labeledcompounds (including deuterium substitutions) and polymorphs of thecompound.

Salts of the compound of formula (I) may be made by methods known to aperson skilled in the art. For example, treatment of a compound offormula (I) with an appropriate base or acid in an appropriate solventwill yield the corresponding salt.

Salts encompassed within the term “pharmaceutically acceptable salts”refer to non-toxic salts of the compound of formula (I). Preferred arealkaline salts of the carboxylic acid, such as sodium, potassium,lithium, calcium, magnesium, aluminium, zinc,N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,ethylenediamine, N-methylglucamine and procaine salts. Other salts,which are not pharmaceutically acceptable, may be useful in thepreparation of compounds of formula (I) and these should be consideredto form a further aspect of the invention.

All starting materials, reagents, acids, bases, solvents and catalystsutilized to synthesize the compounds of formula (I) are eithercommercially available or can be produced by organic synthesis methodsknown to one of ordinary skill in the art. All methods described hereincan be performed in any suitable order unless otherwise indicated hereinor otherwise clearly contradicted by context. The use of any and allexamples, or exemplary language (e.g. such as) provided herein isintended merely to better illuminate the invention and does not pose alimitation on the scope of the invention otherwise claimed.

In one aspect, the present invention provides a method of treating orpreventing infection with hepatitis B virus in a human or animal,comprising administering to the human or animal in need thereof atherapeutically effective amount of a phenylisoxazolylmethylene-naphthalene-ether derivatives having a structure of formula(I), and a pharmaceutically acceptable salt, ester or stereoisomerthereof:

wherein:

R1, R2 and R3 are independently selected from H, halogen, andunsubstituted or halogen substituted C1-6 alkyl and unsubstituted orhalogen substituted C1-6 alkoxy, provided that at least one of R1, R2and R3 is not hydrogen, R0 is selected from unsubstituted or halogensubstituted C1-6 alkyl, C3-6 cycloalkyl, C4-7 alkylcycloalkyl;

X1 and X2 are independently selected from H and halogen;

moiety —O—Z (residue Z linked to the naphthalene ring via an oxygenatom) attaches to the naphthalene ring, wherein Z is a residue selectedfrom 5-10 membered aryl or 5-10 membered heteroaryl optionally havingone or more hetero atoms selected from N, O and S, wherein the 5-10membered aryl or 5-10 membered heteroaryl is substituted by R4 and isoptionally further substituted by R5;

wherein R4 is selected from —COOH, —CH2COOH, —NHSO2CF3, —SO2NH—C1-6alkyl, —SO3H, —CONHSO2-C1-6alkyl, —CONHSO2-C3-6cycloalkyl, —CONHSO2-5-10membered aryl and —CONHSO2-5-10 membered aryl substituted by C1-6 alkylat the aryl, and R5 is selected from H, C1-6 alkyl, halogen, C1-6haloalkyl, —O—(C1-6 alkyl) and —NH—(C1-6 alkyl).

In preferred embodiments of the present invention, R1, R2 and R3 areindependently selected from H, halogen and C1-3 perfluoroalkoxy, such asH, Cl, F and —O—CF3. In one embodiment of the invention, both of R1 andR2 are Cl, and R3 is H. In another embodiment of the invention, both ofR1 and R2 are Cl, and R3 is F. In still another embodiment of theinvention, both of R1 and R2 are Cl, and R3 is —O—CH3. In yet anotherembodiment of the invention, R1 is —O—CF3, and both of R2 and R3 are H.In yet another embodiment of the invention, R0 is isopropyl orcyclopropyl.

In one embodiment of the present invention, Z is a phenyl, which isoptionally substituted by 1-5 halogen atoms. In another embodiment ofthe present invention, Z is a 5-10 membered heteroaryl having one ormore hetero atoms selected from N, O and S. In a preferred embodiment ofthe present invention, Z is a 5-6 membered heteroaryl having one or morehetero atoms selected from N, O and S. In yet another embodiment of thepresent invention, Z is a R4 and optionally R5 substituted pyridyl.

In preferred embodiments of the present invention, R4 is selected from—COOH, —CH2COOH, —CONHSO2-C1-6 alkyl and —CONHSO2-C3-6 cycloalkyl. Inmore preferred embodiment of the present invention. R4 is —COOH or—CH2COOH. In a most preferred embodiment of the present invention, R4 is—COOH.

Preferably, R5 is one selected from H, C1-3 alkyl and halogen.

In a preferred embodiment of the present invention, Z is pyridyl; R4 is—COOH— and R5 is H or halogen.

Preferably, the halogen in the aforesaid substituents is fluoro orchloro.

Specifically, in preferred embodiments of the present invention, thecompound having the formula (I) is of one of the following structures:

The method for preparing the compound of formula (I) can include thefollowing four general routes (Route A, Route B, Route C, and Route D),among which:

Hereinafter, the above-mentioned four general routes will be describedin detail.

Route A:

(a) reacting a halogenated compound of the formula (A1) with adinaphthol to give an ether of the formula (A2). The reaction is carriedout in a polar solvent with a base, preferably, in DMF or acetonitrileor the like with cesium carbonate or potassium carbonate or similarbases.

wherein:

X is a halogen;

R1, R2 and R3 are independently selected from H, halogen, andunsubstituted or halogen substituted C1-6 alkyl and unsubstituted orhalogen substituted C1-6 alkoxy provided that at least one of R1, R2 andR3 is not hydrogen, R0 is selected from unsubstituted or halogensubstituted C1-6 alkyl, C3-6 cycloalkyl, C4-7 alkylcycloalkyl;

(b) reacting the resulting ether of the formula (A2) with a halogenatedcompound X—Z, to give a compound of formula (I),

wherein X is halogen, Z is a residue selected from 5-10 memberedheteroaryl having one or more hetero atoms selected from N, O and S,wherein residue Z is substituted by R4 and optionally furthersubstituted by R5;

wherein R4 is selected from —COOH, —CH2COOH, —NHSO2CF3, —SO2NH—C1-6alkyl, —S03H, —CONHSO2-C1-6 alkyl, —CONHSO2-C3-6 cycloalkyl,—CONHSO2-5-10 membered aryl and —CONHSO2-5-10 membered aryl substitutedby C1-6 alkyl at the aryl, and R5 is selected from H, C1-6 alkyl,halogen and C1-6 haloalkyl; optionally

(c) reacting a compound of the formula (I) containing a —COOHsubstituent with an amide compound to give an amide compound of theformula (I) compound; and optionally

(d) when Z is substituted with R4 selected from —COOH and —CH2COOH,ester precursors can be converted to free acids by hydrolysis usingconditions well known to those skilled in the art,

wherein the compound of the formula (I) is as hereinabove defined.

According to the preparation method as provided by the presentinvention, X is preferably bromine or iodine, and more preferablybromine.

Route B:

(a) reacting a halogenated compound of the formula (A1) with asubstituted naphthol (B1) to give an ether of the formula (B2). Thereaction is carried out in a polar solvent with a base, preferably, inDMF or acetonitrile or the like, with cesium carbonate or potassiumcarbonate or similar bases;

(b) Compound (B2) is converted to boronic ester of the formula (B3),preferably, under Pd-catalyzed conditions;

(c) Compound (B3) is converted to naphthol (A2) by oxidation, withoxidants such as NaClO2 or H2O2;

(d) Compound (A2) is converted to Compound (I) using condition outlinedin Route A,

wherein X3 is a halogen, preferably bromine or iodine, and morepreferably bromine.

Route C:

(a) reacting a substituted naphthol (C1) with halogenated compound X—Zto give an ether of the formula (C2), wherein the reaction is carriedout in a polar solvent with a base, preferably, in DMF or acetonitrileor the like, with cesium carbonate or potassium carbonate or similarbases;

(b) Compound (C2) is converted to boronic ester (C3), preferably, underPd-catalyzed conditions;

(c) Compound (C3) is converted to naphthol (C4) by oxidation, withoxidants such as NaClO2 or H2O2;

(d) Compound (C4) is converted to Compound (I) using similar conditionoutlined in Route A;

wherein X4 is a halogen, preferably bromine or iodine, and morepreferably bromine.

Route D:

reacting a dinaphthol with halogenated compound X—Z to give an ether ofthe formula (C4) using similar condition outlined in Route C:

Compound (C4) is converted to Compound (I) using similar conditionoutlined in Route C.

According to the method of treating or preventing infection withhepatitis B virus in a human or animal, the phenylisoxazolylmethylene-naphthalene-ether derivative having the structure of formula(I), or pharmaceutically acceptable salts, esters or stereoisomersthereof (“compound of formula (I)” for short) can be administered by anyroute appropriate to the condition to be treated. Suitable routesinclude oral, rectal, nasal, topical (including buccal and sublingual),transdermal, vaginal and parenteral (including subcutaneous,intramuscular, intravenous, intradermal, intrathecal and epidural), andthe like. It will be appreciated that the preferred route may vary withfor example the condition of the recipient.

According to the method of the present invention, the compound offormula (I) may be combined with one or more additional therapeuticagents in any dosage amount of the compound of formula (I), with anydose of other therapeutic agents. In one embodiment, the compound offormula (I) is co-administered with other anti-HBV agents, either aspart of the same pharmaceutical composition or in separatepharmaceutical compositions. These agents also can be administered attheir own schedule and through different route. Co-administrationincludes administration of a unit dose of a compound of formula (I)before or after administration of a unit dose of one or more otheranti-HBV agents. In some embodiments, the compound of formula (I) may beadministered within seconds, minutes, or hours of administration of oneor more other anti-HBV agents. In other embodiments, a unit dose of thecompound of formula (I) is administered first, followed byadministration of one or more other anti-HBV agents within seconds,minutes, or hours.

In certain embodiments, when the compound of formula (I) is combinedwith one or more additional therapeutic agents as described herein, thecomponents of the composition are administered as a simultaneous orsequential regimen. When administered sequentially, the combination maybe administered in two or more administrations.

In preferred embodiments of the present invention, the method oftreating or preventing infection with hepatitis B virus in a human oranimal comprises administering thephenylisoxazolylmethylene-naphthalene-ether derivative having thestructure of formula (I), its pharmaceutically acceptable salt, andester or stereoisomer in combination with one or more other anti-HBVagents.

In the above embodiments, wherein the other anti-HBV agents may beselected from antisense oligonucleotide targeting viral mRNA (ASO),short interfering RNAs (siRNA), B- and T-lymphocyte attenuatorinhibitors, CCR2 chemokine antagonist, compounds targeting HBcAg,compounds targeting hepatitis B core antigen (HBcAg), covalently closedcircular DNA (cccDNA) inhibitors, fatty acid synthase inhibitors,cyclophilin inhibitors, cytokines, Endonuclease modulator, epigeneticmodifiers, gene modifiers or editors, HBsAg inhibitors, HBsAg secretionor assembly inhibitors, HBV antibodies, HBV DNA polymerase inhibitors,HBV replication inhibitors, HBV RNAse inhibitors, HBV vaccines, HBVviral entry inhibitors, HBx inhibitors, Hepatitis B structural proteinmodulator, hepatitis B surface antigen (HBsAg) inhibitors, hepatitis Bsurface antigen (HBsAg) secretion or assembly inhibitors, hepatitis Bvirus E antigen inhibitors, hepatitis B virus replication inhibitors,Hepatitis virus structural protein inhibitor, IL-2 agonist, IL-7agonist, immunomodulators, inhibitors of ribonucleotide reductase,Interferon agonist, Interferon alpha 1 ligand, Interferon alpha 2ligand, Interferon alpha 5 ligand modulator, Interferon alpha ligand,Interferon alpha ligand modulator, interferon alpha receptor ligands,Interferon beta ligand, Interferon ligand, Interferon receptormodulator, Interleukin-2 ligand, ipi4 inhibitors, lysine demethylaseinhibitors, microRNA (miRNA) gene therapy agents, modulators of B7-H3,modulators of B7-H4, modulators of CD 160, modulators of CD161,modulators of CD27, modulators of CD47, modulators of CD70, modulatorsof GITR, modulators of TIGIT, modulators of Tim-4, Na+-taurocholatecotransporting polypeptide (NTCP) inhibitors, natural killer cellreceptor 2B4 inhibitors, NOD2 gene stimulator, Nucleoprotein inhibitor,nucleoprotein modulators, PD-1 inhibitors, PD-L1 inhibitors,PEG-Interferon Lambda, Peptidylprolyl isomerase inhibitor, Retinoicacid-inducible gene 1 stimulator, Reverse transcriptase inhibitor,Ribonuclease inhibitor, RNA DNA polymerase inhibitor, short synthetichairpin RNAs (sshRNAs), stimulator of interferon gene (STING) agonists,stimulators of NOD 1, T cell surface glycoprotein CD28 inhibitor, TLR-3agonist, TLR-7 agonist, TLR-8 agonist, TLR-9 agonist, TLR9 genestimulator, toll-like receptor (TLR) modulators, Viral ribonucleotidereductase inhibitor, zinc finger nucleases or synthetic nucleases(TALENs).

In the preferred solution of the present invention, the other anti HBVagents can be selected from HBV vaccines, HBV DNA polymerase inhibitors,immunomodulators, toll-like receptor (TLR) modulators, interferon alphareceptor ligands, hepatitis b surface antigen (HBsAg) inhibitors,cyclophilin inhibitors, HBV viral entry inhibitors, antisenseoligonucleotide targeting viral mRNA, short interfering RNAs (siRNA) andddRNAi endonuclease modulators, ribonucleotide reductase inhibitors, HBVE antigen inhibitors, covalently closed circular DNA (cccDNA)inhibitors, fatty acid synthase inhibitors, HBV antibodies, CCR2chemokine antagonists, retinoic acid-inducible gene 1 stimulators, NOD2stimulators, PD-1 inhibitors, PD-L1 inhibitors, KDM inhibitors, and HBVreplication inhibitors.

In one embodiment, the other anti HBV agent is a nucleot(s)ide. Examplesof HBV DNA polymerase inhibitors include adefovir (HEPSERA*),emtricitabine (EMTRIVA), tenofovir disoproxil fumarate (VIREAD®),tenofovir alafenamide, tenofovir, tenofovir disoproxil, tenofoviralafenamide fumarate, tenofovir alafenamide hemifumarate, tenofovirdipivoxil, tenofovir dipivoxil fumarate, tenofovir octadecyloxyethylester, CMX-157, besifovir, entecavir (BARACLUDE®), entecavir maleate,telbivudine (TYZEKA), pradefovir, devudine, ribavirin, lamivudine (EPIVR-V®), phosphazide, famciclovir, fusolin, metacavir, SNC-0 19754, FMCA,AGX-1009, AR-II-04-26, HIP-1302, tenofovir disoproxil aspartate,tenofovir disoproxil orotate, and HS-10234. Further examples of HBV DNApolymerase inhibitors include filocilovir.

In a particular embodiment, the compound of formula (I) is administeredin combination with entecavir or tenofovir.

In one embodiment, the other anti HBV agent is an immunomodulator.Examples of immunomodulators include TLR agonist R07020531, GS-9620,GS-9688. Examples of immunomodulator also includes PD-1 inhibitors suchas nivolumab, pembrolizumab, pidilizumab, BGB-108, camrelizumab(SHR-1210), PDR-001, PF-06801591, IBI-308, cemiplimab, camrelizumab,sintilimab, tislelizumab (BGB-A317), BCD-100, JNJ-63723283, Zimberelimab(GLS-010, WBP-3055), Balstilimab (AGEN2034), and dostarlimab (TSR-042)

Examples of immunomodulator also include PD-L1 inhibitors such asatezolizumab (RG-7446), avelumab, BGB-A333, BMS-936559 (MDX-1105),durvalumab, CX-072, GX-P2 and envafolimab (KN035, ASC022). PD-L1inhibitors include small molecule inhibitors such as GS-4224,INCB086550.

In a particular embodiment, the compound of formula (I) is administeredin combination with KN035 (ASC022).

In one embodiment, the other anti HBV agent is a HBV vaccine. HBVvaccines include both prophylactic and therapeutic vaccines. Examples ofHBV prophylactic vaccines include Vaxelis, Hexaxim, Heplisav, Mosquirix,DTwP-HBV vaccine, Bio-Hep-B, D/T/P/FlBV7M (LBVP-0101; LBVW-0101),DTwP-Hepb-FIib-IPV vaccine, Heberpenta L, DTwP-HepB-Hib, V-419,CVI-HBV-001, Tetrabhay, hepatitis B prophylactic vaccine (Advax SuperD), Hepatrol-07, GSK-223 192A, ENGERIX B®, recombinant hepatitis Bvaccine (intramuscular, Kangtai Biological Products), recombinanthepatitis B vaccine (Hansenual polymorpha yeast, intramuscular, HualanBiological Engineering), recombinant hepatitis B surface antigenvaccine, Bimmugen, Euforavac, Eutravac, anrix-DTaP-IPV-Hep B, HBAI-20,Infanrix-DT aP-IPV-Hep B-Hib, Pentabio Vaksin DTP-HB-Hib, Comvac 4,Twinrix, Euvax-B, Tritanrix HB, Infanrix Hep B, Comvax, DTP-Hib-HBVvaccine, DTP-HBV vaccine, Yi Tai, Heberbiovac HB, Trivac HB, GerVax,DTwP-Hep B-Hib vaccine, Bilive, HepavaxGene, SUPERVAX, Comvac5,Shanvac-B, Hebsulin, Recombivax HB, Revac B mcf, Revac B+, Fendrix,DTwP-HepB-Hib, DNA-001, Shan5, Shan6, rhHBsAG vaccine, HBI pentavalentvaccine, LBVD, Infanrix HeXa, and DTaP-rHB-Hib vaccine.

In a particular embodiment, the compound of formula (I) is administeredin combination with ABX203, AIC 649, INO-1800, HB-110, TG1050, HepTcell,VR-CHB01, VBI-2601 or CARG-201.

In one embodiment, the other anti HBV agent is antisense oligonucleotidetargeting viral mRNA (ASO). Examples of antisense oligonucleotideinclude Ionis-HBVRx and Ionis-HBV-LRx.

In one embodiment, the other anti HBV agent is short interfering RNAs(siRNA). Examples of short interfering RNAs include JNJ-3989 (ARO-HBV),Vir-2218 (ALN-HBV02), and DCR-HBVS.

In one embodiment, the other anti HBV agent is a hepatitis surfaceantigen (HBsAg) Inhibitor. Examples of HBsAg inhibitors includeHBF-0259, PBHBV-001, PBHBV-2-15, PBHBV-2-1, REP-9AC, REP-9C, REP-9,REP-2139, REP-2139-Ca, REP-2165, REP-2055, REP-2163, REP-2165, REP-2053,REP-203 and REP-006, REP-9 AC, as well as inhibitors targeting the PAPD57.

In one embodiment, the other anti HBV agent is a HB Viral EntryInhibitor, such as Myrcludex B, or an antibody targeting at preS.

In one embodiment, the other anti HBV agent is a fatty acid synthaseinhibitor, such as TVB-2640, TVB-3150, TVB-3199, TVB-3166, TVB-3567 andTVB-3664.

In one embodiment, the other anti HBV agent is a capsid inhibitor.Examples of HBsAg inhibitors include ABI-H0731, ABI-H2158, ABI-H3733,CB-HBV-001, JNJ-6379, JNJ-0440, QL-007, RG-7907 and RO7049389.

In yet another embodiment, the other anti-HBV agent is ordinary orlong-acting interferon. The examples include Pegasys and PEG-INTRON.

In a specific embodiment, the compound of formula (I) of the presentinvention is combined with Pegasys.

In another aspect, the present invention provides use of aphenylisoxazolyl methylene-naphthalene-ether derivative having astructure of formula (I), or a pharmaceutically acceptable salt, esteror stereoisomer thereof in the preparation of a pharmaceuticalcomposition for anti-hepatitis B virus. In this embodiment, thephenylisoxazolyl methylene-naphthalene-ether derivative having thestructure of formula (I), or a pharmaceutically acceptable salt, esteror stereoisomer thereof are as defined above.

According to the use of the present invention, the anti-HBVpharmaceutical composition comprises as an active ingredient aphenylisoxazolyhnethylene-naphthalene-ether derivative having astructure of formula (I), a pharmaceutically acceptable salt, ester orstereoisomer thereof and one or more other anti-HBV agents. In thisembodiment, the other anti-HBV agents are as defined above.

In still another aspect, the present invention provides anpharmaceutical composition for anti-hepatitis B virus comprising atherapeutically effective amount of a phenylisoxazolylmethylene-naphthalene-ether derivative having a structure of formula(I), a pharmaceutically acceptable salt, ester or stereoisomer thereofand one or more other anti-HBV agents, and a pharmaceutically acceptableauxiliary material. In this embodiment, the phenylisoxazolylmethylene-naphthalene-ether derivative having the structure of formula(I), or a pharmaceutically acceptable salt, ester or stereoisomerthereof are as defined above, and the other anti-HBV agents are asdefined above.

The pharmaceutical composition of the present invention are suitable fororal, rectal, nasal, topical (including buccal and sublingual),transdermal, vaginal and parenteral (including subcutaneous,intramuscular, intravenous, intradermal, intrathecal and epidural)administration, and the like, although the most suitable route in anygiven case will depend on the nature and severity of the conditionsbeing treated and on the nature of the active ingredient. Thepharmaceutical compositions may be conveniently presented in unit dosageform and prepared by any of the methods well-known to those skilled inthe art of pharmacy.

The compositions include those suitable for various administrationroutes, including oral administration. The compositions may be presentedin unit dosage form and may be prepared by any of the methods well knownin the art of pharmacy. Such methods include the step of bringing intoassociation the active ingredient (e.g., a compound of the presentdisclosure or a pharmaceutical salt thereof) with one or morepharmaceutically acceptable excipients. The compositions may be preparedby uniformly and intimately bringing into association the activeingredient with liquid excipients or finely divided solid excipients orboth, and then, if necessary, shaping the product. Compositionsdescribed herein that are suitable for oral administration may bepresented as discrete units (a unit dosage form) including but notlimited to capsules, cachets or tablets each containing a predeterminedamount of the active ingredient. In one embodiment, the pharmaceuticalcomposition is a tablet.

The compound may be administered to an individual in an effectiveamount. The amount of active ingredient that may be combined with theinactive ingredients to produce a dosage form may vary depending uponthe intended treatment subject and the particular mode ofadministration. For example, in some embodiments, a dosage form for oraladministration to humans may contain approximately 1 to 1000 mg ofactive material formulated with an appropriate and convenient amount ofa pharmaceutically acceptable excipient. In certain embodiments, thepharmaceutically acceptable excipient varies from about 5 to about 95%of the total compositions.

The dosage or dosing frequency of a compound of the present disclosuremay be adjusted over the course of the treatment, based on the judgmentof the administering physician. The frequency of dosage of the compoundof the present disclosure are will be determined by the needs of theindividual patient and can be, for example, once per day or twice, ormore times, per day. Administration of the compound continues for aslong as necessary to treat the HBV infection.

BRIEF DESCRIPTION OF THE DRAWINGS

In order to explain the embodiments of the present invention or thetechnical solutions in the prior art more clearly, drawings required forthe description of the embodiments of the present invention will bebriefly described below. Obviously, the drawings in the followingdescription are only some embodiments of the present invention.

FIG. 1 is EC50 curves of inhibition HBV DNA in PHH Cells;

FIG. 2 is EC50 curves of HBsAg inhibition in PHH Cells;

FIG. 3 is EC50 curves of HBV RNA inhibition in PHH Cells.

FIG. 4 is HBV DNA level in AAV-HBV mouse serum before and aftertreatment of compound 33.

FIG. 5 is HBV RNA level in AAV-HBV mouse serum before and aftertreatment of compound 33.

FIG. 6 is HBsAg level in AAV-HBV mouse serum before and after treatmentof compound 33.

BEST MODE FOR CARRYING OUT THE INVENTION

The present invention will be further illustrated with reference to theexamples below. It is necessary to state that, the examples below areonly for illustration, but not for limitation of the present invention.Various alterations that are made by a person skilled in the art inaccordance with teaching from the present invention should be within thescope claimed by the claims of the present invention.

Example 1 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinic acid (Compound 1)

(a) Referring to the following reaction equation (Route A), Compound1A-1 (1.0 g, 2.88 mmol, 1 eq.), Compound 1A-2 (0.46 g, 2.88 mmol, 1 eq.)and cesium carbonate (1.88 g, 5.76 mmol, 2 eq.) were dissolved in DMF(10 ml). The reaction was carried out at 65° C. for 2 h. After cooling,10 ml water and 10 ml EA (ethyl acetate) were added for extraction, andthe organic phase was washed with water and concentrated to dryness togive Compound 1A,6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-ol,0.8 g, yield: 65.0%. LCMS (ESI): calculated for C23H17Cl2NO3; [M+H]+:426.1, found: 426.1.

(b) Referring to the following reaction equation, Compound 1A (0.2 g,0.47 mmol, 1 eq.), 6-bromonicotinic acid methyl ester (0.1 g, 0.47 mmol,1 eq.) and cesium carbonate (0.306 g, 0.94 mmol, 2 eq.) were dissolvedin DMF (10 ml). The reaction was carried out at 65° C. for 2 h. Aftercooling, 10 ml water and 10 ml EA were added for extraction, and theorganic phase was washed with water and concentrated to dryness to giveCompound 1B, methyl6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalene-2-yl)oxy)nicotinate,0.21 g, yield: 80.0%. LCMS (ESI): calculated for C30H22Cl2N2O5; [M+H]+:561.1, found: 561.1.

(c) Referring to the following reaction equation, compound 1B (100 mg)was dissolved in methanol (2 ml), then 10% NaOH aqueous solution (1 ml)was added, the temperature was raised to 60° C., and the reaction wascarried out for 1 h. The pH of the reaction solution was adjusted to 2to 4 by adding 1N HCl solution, and 10 ml EA (ethyl acetate) was addedfor extraction. The organic phase was concentrated and purified on acolumn (PE/EA/AcOH=1/1/0.01 elution, wherein PE is petroleum ether) togive the title compound 1 (36 mg, yield: 37.0%).

1H NMR (400 MHz, DMSO-d6) δ 8.57 (s, 1H), 8.23 (d, J=7.2 Hz, 1H), 7.74(dd, J=2.0, 8.8 Hz, 2H), 7.60 (d, J=7.6 Hz, 2H), 7.56 (s, 1H), 7.51 (dd,J=8.8, 7.2 Hz, 1H), 7.33 (s, 1H), 7.26 (d, J=8.8 Hz, 1H), 7.02 (d, J=8.0Hz, 1H), 6.93 (d, J=6.4 Hz, 1H), 4.98 (s, 2H), 2.57-2.50 (m, 1H),1.19-1.11 (m, 4H). LCMS (ESI): calculated for C29H20Cl2N2O5; [M+H]+:547.1, found: 547.1, 13C NMR (400 MHz, DMSO-d6) δ 7.79, 8.87, 8.87,59.31, 107.74, 110.05, 110.97, 117.64, 119.43, 122.52, 127.55, 128.64,128.89, 128.89, 129.18, 129.67, 131.73, 131.79, 132.94, 135.10, 135.10,141.20, 149.11, 150.73, 155.79, 159.68, 163.82, 167.81, 172.61. IR(cm-1): major stretches at 1591.94 (C═O stretch), 1412.27, 1556.70 (C—Cstretch), 1364.37, 1389.89 (C—H deformation), 1218.41, 1250.94 (C═Nstretch), 791.88 (C—Cl stretch).

Example 2 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)pyridazine-3-carboxylic acid (Compound 2)

Following the procedure of Example 1, the title Compound 2 was obtainedby substituting methyl 6-bromopyridazine-3-carboxylate for6-bromonicotinic acid methyl ester.

1H NMR (400 MHz, DMSO-d6) δ 8.52 (s, 1H), 8.25 (d, J=7.2 Hz, 1H), 7.74(dd, J=2.0, 8.8 Hz, 2H), 7.61 (d, J=7.6 Hz, 2H), 7.52 (dd, J=8.8, 7.2Hz, 1H), 7.34 (s, 1H), 7.26 (d, J=8.8 Hz, 1H), 7.00 (d, J=8.0 Hz, 1H),6.95 (d, J=6.4 Hz, 1H), 4.98 (s, 2H), 2.59-2.50 (m, 1H), 1.21-1.11 (m,4H). LCMS (ESI): calculated for C28H19Cl2N3O5; [M+H]+: 548.1, found:548.1.

Example 3 Preparation of5-chloro-6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinic acid (Compound 3)

Following the procedure of Example 1, the title Compound 3 was obtainedby substituting methyl 5,6-dichloronicotinate for 6-bromonicotinic acidmethyl ester.

1H NMR (400 MHz, DMSO-d6) δ 8.60 (s, 1H), 7.73 (dd, J=2.0, 8.8 Hz, 2H),7.59 (d, J=7.6 Hz, 2H), 7.51 (dd, J=8.8, 7.2 Hz, 1H), 7.33 (s, 1H), 7.26(d, J=8.8 Hz, 1H), 7.01 (d, J=8.0 Hz, 1H), 6.95 (d, J=6.4 Hz, 1H), 5.00(s, 2H), 1.26-1.12 (m, 511). LCMS (ESI): calculated for C29H19C3N2O5;[M+H]+: 581.0, found: 581.0.

Example 4 Preparation of2-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)thiazole-5-carboxylic acid (Compound 4)

Following the procedure of Example 1, the title Compound 4 was obtainedby substituting methyl 2-bromothiazole-5-carboxylate for6-bromonicotinic acid methyl ester.

1H NMR (400 MHz, DMSO-d6) δ 8.80 (s, 1H), 7.69 (dd, J=2.0, 8.8 Hz, 2H),7.59 (d, J=7.6 Hz, 2H), 7.53 (dd, J=8.8, 7.2 Hz, 1H), 7.32 (s, 1H), 7.26(d, J=8.8 Hz, 1H), 7.01 (d, J=8.0 Hz, 1H), 6.99 (d, J=6.4 Hz, 1H), 5.00(s, 2H), 1.25-1.12 (m, 5H). LCMS (ESI): calculated for C27H18Cl2N2O5S;[M+H]+: 553.0, found: 553.0.

Example 5 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)-5-methylnicotinic acid (Compound 5)

Following the procedure of Example 1, the title Compound 5 was obtainedby substituting methyl 6-bromo-5-methylnicotinate for 6-bromonicotinicacid methyl ester.

1H NMR (400 MHz, DMSO-d6) δ 12.78 (s, 1H), 8.35 (d, J=1.5 Hz, 1H),8.12-7.90 (m, 1H), 7.72-7.61 (m, 2H), 7.54 (s, 3H), 7.28 (m, 2H),7.15-7.10 (m, 1H), 7.07 (dd, J=7.5, 1.5 Hz, 1H), 6.95 (dd, J=7.6, 1.6Hz, 1H), 5.41 (s, 2H), 2.99-2.70 (m, 1H), 2.28 (s, 3H), 2.12-1.56 (m,4H). LCMS (ESI): calculated for C30H22Cl2N2O5; [M+H]+: 561.1, found:561.1.

Example 6 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)-N-(cyclopropylsulfonyl)nicotinamide(Compound 6)

Compound 1 (70 mg) as prepared in Example 1 and cyclopropylsulfonamide(23 mg) were dissolved in 2 ml DCM (dichloromethane), then 40 mg EDCI(1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) and 26 mgDMAP (dimethylaminopyridine) were added. After completion of thereaction, 10 ml DCM and 10 ml water was added for extraction. Theorganic phase was washed with water and concentrated to dryness. Thecrude product is purified by column (PE/EA/AcOH=2/1/0.01) to give thetitle Compound 6 (8 mg, yield: 9.6%).

1H NMR (400 MHz, DMSO-d6) δ 8.63 (d, J=1.5 Hz, 1H), 8.30 (dd, J=7.5, 1.5Hz, 1H), 7.79 (d, J=8.0 Hz, 1H), 7.73 (d, J=8.0 Hz, 1H), 7.59-7.62 (m,3H), 7.49-7.53 (m, 1H), 7.35 (s, 1H), 7.26-7.29 (m, 1H), 7.10 (d, J=8.0Hz, 1H), 6.93-6.96 (m, 1H), 4.98 (s, 2H), 1.02-1.20 (m, 10H). LCMS(ESI): calculated for C32H25Cl2N3O6S; [M+H]+: 650.1, found: 650.1.

Example 7 Preparation of5-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)pyrazine-2-carboxylic acid (Compound 7)

Following the procedure of Example 1, the title Compound 7 was obtainedby substituting methyl 5-chloro-pyridine-2-carboxylate for6-bromonicotinic acid methyl ester.

1H NMR (400 MHz, DMSO-d6) δ 8.63 (s, 1H), 8.30 (s, 1H), 7.79 (d, J=8.8Hz, 1H), 7.72 (d, J=9.2 Hz, 1H), 7.58-7.63 (m, 4H), 7.49-7.53 (m, 1H),7.34 (d, J=2.0 Hz, 1H), 6.94 (d, J=9.2 Hz, 1H), 4.98 (s, 2H), 1.11-1.22(m, 5H). LCMS (ESI): calculated for C28H19Cl2N3O5; [M+H]+: 548.1, found:548.1.

Example 8 Preparation of2-chloro-6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinic acid (Compound 8)

Following the procedure of Example 1, the title Compound 8 was obtainedby substituting methyl 2,6-dichloronicotinate for 6-bromonicotinic acidmethyl ester.

1H NMR (400 MHz, DMSO-d6) δ 7.98 (br s, 1H), 7.70-7.79 (m, 2H), 7.60 (d,J=8.0 Hz, 2H), 7.47-7.55 (m, 211), 7.18-7.33 (m, 2H), 6.90-6.95 (m, 2H),4.98 (s, 2H), 1.11-1.22 (m, 5H). LCMS (ESI): calculated forC29H19Cl3N2O5; [M+H]+: 581.0, found: 581.0.

Example 9 Preparation of5-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)picolinicacid (Compound 9)

Following the procedure of Example 1, the title Compound 9 was obtainedby substituting methyl 5-bromopicolinate for 6-bromonicotinic acidmethyl ester.

1H NMR (400 MHz, DMSO-d6) δ 8.46 (d, J=3.1 Hz, 1H), 8.03 (d, J=8.7 Hz,11H), 7.83 (d, J=8.9 Hz, 1H), 7.72 (d, J=9.1 Hz, 1H), 7.55 (dt, J=28.7,8.3 Hz, 411), 7.43 (d, J=8.6 Hz, 1H), 7.39-7.24 (m, 2H), 6.95 (d, J=8.9Hz, 1H), 4.98 (s, 2H), 1.23-1.02 (m, 5H). LCMS (ESI): calculated forC29H20Cl2N2O5; [M+H]+: 547.1, found: 547.1.

Example 10 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)-2-methylnicotinicacid (Compound 10)

Following the procedure of Example 1, the title Compound 10 was obtainedby substituting methyl 6-chloro-2-methylnicotinate for 6-bromonicotinicacid methyl ester.

1H NMR (400 MHz, DMSO-d6) δ 8.21 (d, J=8.6 Hz, 1H), 7.78 (d, J=8.9 Hz,1H), 7.72 (d, J=9.0 Hz, 1H), 7.62-7.55 (m, 3H), 7.51 (dd, J=9.0, 7.1 Hz,1H), 7.34 (d, J=2.5 Hz, 1H), 7.27 (dd, J=8.8, 2.4 Hz, 1H), 6.94 (dd,J=8.9, 2.5 Hz, 1H), 6.85 (d, J=8.6 Hz, 1H), 4.98 (s, 2H), 2.52 (s, 3H),1.24-1.07 (m, 5H). LCMS (ESI): calculated for C30H22Cl2N2O5; [M+H]+:561.1, found: 561.1.

Example 11 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)picolinicacid (Compound 11)

Following the procedure of Example 1, the title Compound 11 was obtainedby substituting methyl 2,6-dichloronicotinate for 6-bromonicotinic acidmethyl ester.

1H NMR (400 MHz, DMSO-d6) δ 7.99 (t, J=7.9 Hz, 1H), 7.77 (d, J=8.1 Hz,2H), 7.71 (d, J=8.9 Hz, 1H), 7.63-7.45 (m, 4H), 7.33 (s, 1H), 7.29 (d,J=9.0 Hz, 1H), 7.22 (d, J=8.2 Hz, 1H), 6.94 (d, J=9.0 Hz, 1H), 4.98 (s,2H), 1.26-1.01 (m, 5H). LCMS (ESI): calculated for C29H20Cl2N2O5;[M+H]+: 547.1, found: 547.1.

Example 12 Preparation of2-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)isonicotinicacid (Compound 12)

Following the procedure of Example 1, the title Compound 12 was obtainedby substituting methyl 2-fluoroisonicotinate for 6-bromonicotinic acidmethyl ester.

1H NMR (400 MHz, DMSO-d6) δ 8.29 (d, J=5.1 Hz, 1H), 7.78 (d, J=8.8 Hz,1H), 7.59 (t, J=7.7 Hz, 3H), 7.52 (m, 2H), 7.34 (s, 2H), 7.29 (s, 1H),4.98 (s, 2H), 1.31-1.06 (m, 5H). LCMS (ESI): calculated forC29H20Cl2N2O5; [M+H]+: 547.1, found: 547.1.

Example 13 Preparation of3-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)picolinicacid (Compound 13)

Following the procedure of Example 1, the title Compound 13 was obtainedby substituting methyl 3-fluoropicolinate for 6-bromonicotinic acidmethyl ester.

1H NMR (400 MHz, DMSO-d6) δ 8.39 (d, J=4.4 Hz, 1H), 7.77 (d, J=9.0 Hz,1H), 7.66 (d, J=9.2 Hz, 1H), 7.59 (d, J=8.1 Hz, 2H), 7.54-7.43 (m, 3H),7.30 (d, J=2.8 Hz, 2H), 7.26-7.16 (m, 1H), 6.95-6.85 (m, 1H), 4.95 (s,2H), 1.24-1.06 (m, 5H). LCMS (ESI): calculated for C29H20Cl2N2O5;[M+H]+: 547.1, found: 547.1.

Example 14 Preparation of2-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)benzoicacid (Compound 14)

Following the procedure of Example 1, the title Compound 14 was obtainedby substituting methyl 2-fluorobenzoate for 6-bromonicotinic acid methylester.

1H NMR (400 MHz, DMSO-d6) δ 7.82 (d, J=7.8 Hz, 1H), 7.73 (d, J=8.9 Hz,1H), 7.66-7.57 (m, 3H), 7.55-7.45 (m, 2H), 7.26 (d, J=10.6 Hz, 2H),7.21-7.11 (m, 2H), 6.99 (d, J=8.3 Hz, 1H), 4.94 (s, 2H), 1.27-1.06 (m,5H). LCMS (ESI): calculated for C30H21Cl2NO5; [M+H]+: 546.1, found:546.1

Example 15 Preparation of2-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)nicotinicacid (Compound 15

Following the procedure of Example 1, the title Compound 15 was obtainedby substituting methyl 2-chloronicotinate for 6-bromonicotinic acidmethyl ester.

1H NMR (400 MHz, DMSO-d6) δ 8.30-8.19 (m, 2H), 7.73 (dd, J=19.2, 9.0 Hz,2H), 7.60 (d, J=7.9 Hz, 2H), 7.55-7.47 (m, 2H), 7.32 (d, J=2.5 Hz, 1H),7.26-7.18 (m, 2H), 6.92 (dd, J=8.9, 2.5 Hz, 1H), 4.98 (s, 2H), 1.23-1.10(m, 5H). LCMS (ESI): calculated for C29H20Cl2N2O5; [M+H]+: 547.1, found:547.1.

Example 16 Preparation of3-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)isonicotinicacid (Compound 16)

Following the procedure of Example 1, the title Compound 16 was obtainedby substituting methyl 3-fluoroisonicotinate for 6-bromonicotinic acidmethyl ester.

1H NMR (400 MHz, DMSO-d6) δ 8.52 (d, J=4.8 Hz, 1H), 8.39 (s, 1H), 7.76(d, J=8.9 Hz, 1H), 7.73 (d, J=4.9 Hz, 1H), 7.64 (d, J=9.0 Hz, 1H), 7.59(d, J=7.7 Hz, 2H), 7.50 (dd, J=9.0, 7.0 Hz, 1H), 7.32-7.15 (m, 4H), 6.89(dd, J=8.9, 2.5 Hz, 1H), 4.95 (s, 2H), 1.27-1.09 (m, 5H). LCMS (ESI):calculated for C29H20Cl2N2O5; [M+H]+: 547.1, found: 547.1.

Example 17 Preparation of6-((6-((5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinic acid (Compound 17)

Following the procedure of Example 1, the title Compound 17 was obtainedby substituting4-(chloromethyl)-5-cyclopropyl-3-(2-(trifluoromethoxy)phenyl) isoxazolefor 1A-1.

1H NMR (400 MHz, DMSO-d6) δ 8.62 (s, 1H), 8.27 (d, J=8.1 Hz, 1H), 7.75(dt, J=31.9, 15.9 Hz, 2H), 7.61 (s, 2H), 7.56-7.43 (m, 2H), 7.36 (s,1H), 7.29 (d, J=8.6 Hz, 1H), 7.11 (d, J=8.2 Hz, 1H), 7.00 (d, J=8.5 Hz,1H), 5.03 (s, 211), 2.44-2.37 (m, 1H), 1.20-1.05 (m, 4H). LCMS (ESI):calculated for C30H21F3N206; [M+H]+: 563.1, found: 563.1.

Example 18 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichloro-4-fluorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinic acid (Compound 18)

Following the procedure of Example 1, the title Compound 18 was obtainedby substituting4-(chloromethyl)-5-cyclopropyl-3-(2,6-dichloro-4-fluorophenyl) isoxazolefor 1A-1.

1H NMR (400 MHz, DMSO-d6) δ 8.63 (s, 1H), 8.27 (d, J=7.9 Hz, 1H),7.87-7.63 (m, 4H), 7.60 (s, 1H), 7.40-7.24 (m, 2H), 7.11 (d, J=8.0 Hz,1H), 6.96 (d, J=8.6 Hz, 1H), 4.98 (s, 2H), 2.47-2.40 (m, 1H), 1.23-1.08(m, 4H). LCMS (ESI): calculated for C29H19Cl2FN2O5; [M+H]+: 565.1,found: 565.1.

Example 19 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichloro-4-methoxyphenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinic acid (Compound 19)

Following the procedure of Example 1, the title Compound 19 was obtainedby substituting4-(chloromethyl)-5-cyclopropyl-3-(2,6-dichloro-4-methoxyphenyl)isoxazole for 1A-1.

LCMS (ESI): calculated for C30H22Cl2N206; [M+H]+: 577.1, found: 577.1.

Example 20 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-1-fluoronaphthalen-2-yl)oxy)nicotinicacid (Compound 20)

(a) Referring to the following reaction equation (Route C), Compound20A-1 (1.0 g, 4.15 mmol, 1 eq.), Compound 20A-2 (0.90 g, 4.15 mmol, 1eq.) and cesium carbonate (2.70 g, 8.30 mmol, 2 eq.) were dissolved inDMF (10 ml). The reaction was carried out at 65° C. for 2 h. Aftercooling, 10 ml water and 10 ml EA (ethyl acetate) were added forextraction, and the organic phase was washed with water and concentratedto dryness to give Compound 20A, methyl6-((6-bromo-1-fluoronaphthalen-2-yl)oxy)nicotinate, 1.2 g, yield: 77.0%.LCMS (ESI): calculated for C17H11BrFNO3; [M+H]+: 376.0, found: 376.0.

(b) Referring to the following reaction equation, compound 20A (200 mg,0.53 mmol, 1 eq) was dissolved in dry THF (2 ml), then KOAc (104 mg,1.06 mmol, 2 eq), Pd(dppf)2C12 (39 mg, 0.053 mmol, 0.1 eq), andbis(pinacolato)diboron (135 mg, 0.53 mmol, 1 eq) were added under N2,and the reaction mixture was heated to reflux for 2 h. After cooling, 10ml water and 10 ml EtOAc were added for extraction, and the organicphase was washed with water and concentrated to dryness. The residue waspurified by silica gel column chromatography (petroleum Ether:EtOAc=3:1)to give Compound 20B, methyl6-((1-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)oxy)nicotinate,151 mg, yield: 67.1%. LCMS (ESI): calculated for C23H23BFNO5; [M+H]+:424.2, found: 424.2.

(c) Referring to the following reaction equation, compound 20B (100 mg)was dissolved in EtOH (2 ml), then 30% H₂O₂ aqueous solution (1 ml) wereadded. The reaction mixture was stirred at room temperature for 1 h,quenched with saturated aqueous Na2SO3, and extracted with EA. Theorganic phase was concentrated and purified on a column (PE/EA=3/1) togive the compound 20C (36 mg, yield: 37.0%). LCMS (ESI): calculated forC17H12FNO4; [M+H]+: 314.1, found: 314.1.

(d) Referring to the following reaction equation, Compound 20C (0.2 g,0.47 mmol, 1 eq.), 1A-1 (0.1 g, 0.47 mmol, 1 eq.) and cesium carbonate(0.306 g, 0.94 mmol, 2 eq.) were dissolved in DMF (10 ml) for reacting.The reaction was carried out at 65° C. for 2 h. After cooling, 10 mlwater and 10 ml EtOAc were added for extraction, and the organic phasewas washed with water and concentrated to dryness to give Compound 20D,0.21 g, yield: 80.0%. LCMS (ESI): calculated for C30H21Cl2FN2O5; [M+H]+:579.1, found: 579.1.

(e) Referring to the following reaction equation, compound 20D (100 mg)was dissolved in dry THF (2 ml), then 10% NaOH aqueous solution (1 ml)were added under N2, and the reaction mixture was heated to reflux for 1h. The pH of the reaction solution was adjusted to 3 to 4 by adding 1NHCl solution, and 10 ml EA was added for extraction. The organic phasewas concentrated and purified on a column (PE/EA/AcOH=1/1/0.01 elution)to give the title compound 20 (36 mg, yield: 37.0%).

1H NMR (400 MHz, DMSO-d6) δ 8.63 (d, J=2.4 Hz, 1H), 8.30 (dd, J=8.7, 2.4Hz, 1H), 7.92 (d, J=9.0 Hz, 1H), 7.70 (s, 1H), 7.64 (d, J=5.3 Hz, 1H),7.60 (d, J=6.4 Hz, 1H), 7.57 (d, J=4.3 Hz, 2H), 7.42-7.36 (m, 2H), 7.17(d, J=8.6 Hz, 1H), 5.09 (s, 2H), 1.22-1.06 (m, 5H). LCMS (ESI):calculated for C29H19Cl2FN2O5; [M+H]+: 565.1, found: 565.1.

Example 21 Preparation of6-((1-chloro-6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinic acid (Compound 21)

Following the procedure of Example 20, the title Compound 21 wasobtained by substituting 6-bromo-1-chloronaphthalen-2-ol for 20A-1.

1H NMR (400 MHz, DMSO-d6) δ 8.63 (s, 1H), 8.27 (d, J=7.9 Hz, 1H),7.87-7.63 (m, 411), 7.60 (s, 1H), 7.40-7.24 (m, 211), 7.11 (d, J=8.0 Hz,1H), 6.96 (d, J=8.6 Hz, 1H), 4.98 (s, 2H), 2.47-2.40 (m, 1H), 1.23-1.08(m, 4H). LCMS (ESI): calculated for C29H19Cl3N2O5; [M+H]+: 581.0, found:581.0.

Example 22 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)nicotinicacid (Compound 22)

(a) Referring to the following reaction equation (Route D), Compound22A-1 (2.0 g, 12.49 mmol, 1 eq.), Compound 22A-2 (1.71 g, 9.99 mmol, 0.8eq.) and cesium carbonate (6.09 g, 18.74 mmol, 1.5 eq.) were dissolvedin DMF (20 ml) for reacting. The reaction was carried out at 65° C. for3 h. After cooling, 30 ml water and 30 ml EA (ethyl acetate) were addedfor extraction, and the organic phase was washed with water andconcentrated to dryness. The residue was purified by silica gel columnchromatography (petroleum:AcOEt=5:1) to give Compound 22A, methyl6-((6-hydroxynaphthalen-1-yl)oxy)nicotinate, 1.1 g, yield: 37.3%. LCMS(ESI): calculated for C17H13NO4; [M+H]+: 296.1, found: 296.1.

(b) Referring to the following reaction equation, Compound 22A (0.2 g,0.68 mmol, 1 eq.), 22A-3 (0.2 g, 0.68 mmol, 1 eq.) and cesium carbonate(0.44 g, 1.36 mmol, 2 eq.) were dissolved in DMF (5 ml) for reacting.The reaction was carried out at 40° C. for 2 h. After cooling, 10 mlwater and 10 ml EA were added for extraction, and the organic phase waswashed with water and concentrated to dryness to give Compound 22B,methyl6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)nicotinate, 0.31 g, yield: 81.2%. LCMS (ESI): calculated forC30H22Cl2N2O5; [M+H]+: 561.1, found: 561.1.

(c) Referring to the following reaction equation, compound 22B (100 mg)was dissolved in methanol (2 ml), then 10% NaOH aqueous solution (1 ml)was added, the temperature was raised to 60° C., and the reaction wascarried out for 0.5 h. The pH of the reaction solution was adjusted to 2to 4 by adding 1N HCl solution, and 10 ml EA was added for extraction.The organic phase was concentrated on a column (PE/EA/AcOH=1/1/0.01elution) to give the title compound 22 (42 mg, yield: 43.2%).

1H NMR (400 MHz, DMSO-d6) δ 13.11 (br s, 1H), 8.56 (s, 1H), 8.28 (d,J=8.5 Hz, 1H), 7.66 (d. J=8.3 Hz, 1H), 7.56-7.61 (m, 3H), 7.45-7.53 (m,2H), 7.39 (s, 1H), 7.15 (t, J=9.6 Hz, 2H), 6.9 (d, J=9.2 Hz, 2H), 4.98(s, 2H), 1.09-1.28 (m, 5H). LCMS (ESI): calculated for C29H20Cl2N2O5;[M+H]+: 547.1, found: 547.1.

Example 23 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)picolinicacid (Compound 23)

Following the procedure of Example 22, the title Compound 23 wasobtained by substituting methyl 6-fluoropicolinate for 22A-2.

1H NMR (400 MHz, DMSO-d6) δ7.99 (t, J=7.8 Hz, 1H), 7.78 (d, J=7.4 Hz,1H), 7.73 (d, J=9.2 Hz, 1H), 7.63 (d, J=8.3 Hz, 1H), 7.60-7.55 (m, 2H),7.52-7.44 (m, 2H), 7.40-7.37 (m, 1H), 7.20 (d, J=8.3 Hz, 1H), 7.09 (d,J=7.5 Hz, 1H), 6.94-6.90 (m, 1H), 4.99 (s, 2H), 1.23-1.09 (m, 5H). LCMS(ESI): calculated for C29H20Cl2N2O5; [M+H]+: 547.1, found: 547.1.

Example 24 Preparation of2-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)isonicotinicacid (Compound 24)

Following the procedure of Example 22, the title Compound 24 wasobtained by substituting methyl 2-fluoroisonicotinate for 22A-2.

1H NMR (400 MHz, DMSO-d6) δ 8.22 (d, J=5.1 Hz, 1H), 7.67-7.62 (m, 2H),7.58 (d, J=8.0 Hz, 2H), 7.53-7.42 (m, 3H), 7.38 (s, 2H), 7.11 (d, J=7.5Hz, 1H), 6.89 (dd, J=9.2, 2.4 Hz, 1H), 4.98 (s, 2H), 1.22-1.07 (m, 5H).LCMS (ESI): calculated for C29H20Cl2N2O5; [M4+H]+: 547.1, found: 547.1.

Example 25 Preparation of3-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)picolinicacid (Compound 25)

Following the procedure of Example 22, the title Compound 25 wasobtained by substituting methyl 3-fluoropicolinate for 22A-2.

1H NMR (400 MHz, DMSO-d6) δ 8.41-8.37 (m, 1H), 7.90 (d, J=9.2 Hz, 1H),7.62-7.54 (m, 3H), 7.53-7.47 (m, 2H), 7.43-7.35 (m, 2H), 7.35-7.30 (m,1H), 6.99-6.94 (m, 1H), 6.77 (d, J=7.6 Hz, 1H), 5.00 (s, 2H), 1.21-1.10(m, 5H). LCMS (ESI): calculated for C29H20Cl2N2O5; [M+H]+: 547.1, found:547.1.

Example 26 Preparation of2-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)-4-fluorobenzoicacid (Compound 26)

Following the procedure of Example 22, the title Compound 26 wasobtained by substituting methyl 2,4-difluorobenzoate for 22A-2.

1H NMR (400 MHz, DMSO-d6) δ 8.22 (d, J=5.1 Hz, 1H), 7.67-7.61 (m, 2H),7.57 (s, 2H), 7.52-7.43 (m, 3H), 7.38 (s, 2H), 7.11 (d, J=7.5 Hz, 1H),6.90 (s, OH), 4.98 (s, 2H), 1.20-1.06 (m, 5H). LCMS (ESI): calculatedfor C30H20Cl2FNO5; [M+H]+: 564.1, found: 564.1.

Example 27 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)-2-methylnicotinicacid (Compound 27)

Following the procedure of Example 22, the title Compound 27 wasobtained by substituting methyl 6-chloro-2-methylnicotinate for 22A-2.

1H NMR (400 MHz, DMSO-d6) δ 8.20 (d, J=8.5 Hz, 1H), 7.63 (dd, J=8.8, 4.5Hz, 2H), 7.57 (s, 1H), 7.52-7.43 (m, 1H), 7.38 (d, J=2.7 Hz, 1H), 7.11(d, J=7.5 Hz, 1H), 6.90 (dd, J=9.2, 2.5 Hz, 1H), 6.81 (d, J=8.6 Hz, 1H),4.98 (s, 2H), 2.48 (s, 3H), 1.23-1.00 (m, 5H). LCMS (ESI): calculatedfor C30H22Cl2N2O5; [M+H]+: 561.1, found: 561.1.

Example 28 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)-5-methylnicotinicacid (Compound 28)

Following the procedure of Example 22, the title Compound 28 wasobtained by substituting methyl 6-chloro-5-methylnicotinate for 22A-2.

1H NMR (400 MHz, DMSO-d6) δ 8.31 (d, J=2.3 Hz, 1H), 8.19 (d, J=2.3 Hz,1H), 7.64 (d, J=8.3 Hz, 1H), 7.61-7.55 (m, 3H), 7.53-7.44 (m, 2H), 7.38(d, J=2.7 Hz, 1H), 7.11 (d, J=7.5 Hz, 1H), 6.88 (dd, J=9.1, 2.5 Hz, 1H),4.98 (s, 2H), 2.47 (s, 3H), 1.20-1.08 (m, 5H). LCMS (ESI): calculatedfor C30H22Cl2N2O5; [M+H]+: 561.1, found: 561.1.

Example 29 Preparation of6-((5-chloro-6-((5-cyclopropyl-3-(2,6-dichlorophenyl)-isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinicacid (Compound 29)

Following the procedure of Example 32, the title Compound 29 wasobtained by substituting 6-bromo-1-chloronaphthalen-2-ol for 32A-1.

1H NMR (400 MHz, DMSO-d6) δ 8.63 (s, 1H), 8.29 (d, J=8.6 Hz, 1H), 7.92(d, J=8.9 Hz, 1H), 7.70 (s, 1H), 7.65-7.46 (m, 4H), 7.38 (s, 2H), 7.17(d, J=8.5 Hz, 1H), 5.09 (s, 2H), 1.21-1.02 (m, 5H). LCMS (ESI):calculated for C29H19Cl3N2O5; [M+H]+: 581.0, found: 581.0.

Example 30 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-2-fluoronaphthalen-1-yl)oxy)nicotinicacid (Compound 30)

Following the procedure of Example 20, the title Compound 30 wasobtained by substituting 6-bromo-2-fluoronaphthalen-1-ol for 20A-1.

1H NMR (400 MHz, DMSO-d6) δ 8.59 (d, J=2.4 Hz, 1H), 8.31 (dd, J=8.6, 2.4Hz, 1H), 7.86 (d. J=9.1 Hz, 1H), 7.66 (d, J=6.9 Hz, 1H), 7.59 (s, 1H),7.56 (dd, J=5.7, 3.3 Hz, 1H), 7.51 (dd, J=9.0, 7.1 Hz, 1H), 7.46-7.39(m, 2H), 7.25 (d, J=8.6 Hz, 1H), 7.05 (dd, J=9.2, 2.4 Hz, 1H), 5.02 (s,2H), 1.28-1.08 (m, 5H). LCMS (ESI): calculated for C29H19Cl2FN2O5;[M+H]+: 565.1, found: 565.1.

Example 31 Preparation of6-((7-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinicacid (Compound 31)

Following the procedure of Example 1, the title Compound 31 was obtainedby substituting naphthalene-2,7-diol for 1A-2.

1H NMR (400 MHz, DMSO-d6) δ 8.64 (s, 1H1), 8.28 (d, J=8.6 Hz, 1H), 7.85(d, J=8.8 Hz, 1H), 7.77 (d, J=9.0 Hz, 1H), 7.58 (d, J=8.1 Hz, 2H),7.55-7.43 (m, 2H), 7.27 (s, 1H), 7.13 (t, J=9.9 Hz, 2H), 6.89 (d, J=8.9Hz, 1H), 4.95 (s, 2H), 1.29-1.06 (m, 5H). LCMS (ESI): calculated forC29H20Cl2N2O5; [M+H]+: 547.1, found: 547.1.

Example 32 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-5-fluoronaphthalen-2-yl)oxy)nicotinicacid (Compound 32)

(a) Referring to the following reaction equation (Route B), Compound32A-1 (1.0 g, 4.15 mmol, 1 eq.), Compound 1A-1 (1.44 g, 4.15 mmol, 1eq.) and cesium carbonate (2.70 g, 8.30 mmol, 2 eq.) were dissolved inDMF (10 ml) for reacting. The reaction was carried out at 65° C. for 2h. After cooling, 10 ml water and 10 ml EA (ethyl acetate) were addedfor extraction, and the organic phase was washed with water andconcentrated to dryness to give Compound 32A,4-(((6-bromo-1-fluoronaphthalen-2-yl)oxy)methyl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole,1.51 g, yield: 71.9%. LCMS (ESI): calculated for C23H15BrCl2FNO2;[M+H]+: 506.0, found: 506.0.

(b) Referring to the following reaction equation, compound 32A (200 mg,0.39 mmol, 1 eq) was dissolved in dry THF (2 ml), then KOAc (76 mg, 0.78mmol, 2 eq), Pd(dppf)2Cl2 (28 mg, 0.039 mmol, 0.1 eq), andbis(pinacolato)diboron (100 mg, 0.39 mmol, 1 eq) were added under N2,and the reaction mixture was heated to reflux for 2 h. After cooling, 10ml water and 10 ml EA were added for extraction, and the organic phasewas washed with water and concentrated to dryness. The residue waspurified by silica gel column chromatography (petroleum:AcOEt=3:1) togive Compound 32B,5-cyclopropyl-3-(2,6-dichlorophenyl)-4-(((1-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-yl)oxy)methyl)isoxazole,137 mg, yield: 62.8%. LCMS (ESI): calculated for C29H27BCl2FNO4; [M+H]+:554.1, found: 554.1.

(c) Referring to the following reaction equation, compound 32B (100 mg)was dissolved in EtOH (2 ml), then 30% H₂O₂ aqueous solution (1 ml) wereadded. The reaction mixture was stirred at room temperature for 1 h,quenched with saturated aqueous Na2SO3, and extracted with EA. Theorganic phase was concentrated and purified on a column (PE/EA=3/1) togive the compound 32C (61 mg, yield: 76.2%). LCMS (ESI): calculated forC23H16Cl2FNO3; [M+H]+: 444.1, found: 444.1.

(d) Referring to the following reaction equation, Compound 32C (50 mg,0.11 mmol, 1 eq.), 1A-3 (24.3 mg, 0.11 mmol, 1 eq.) and cesium carbonate(71.5 mg, 0.22 mmol, 2 eq.) were dissolved in DMF (1 ml) for reacting.The reaction was carried out at 65° C. for 2 h. After cooling, 5 mlwater and 5 ml EA were added for extraction, and the organic phase waswashed with water and concentrated to dryness to give Compound 32D, 40mg, yield: 61.5%. LCMS (ESI): calculated for C30H21Cl2FN2O5; [M+H]+:579.1, found: 579.1.

(e) Referring to the following reaction equation, compound 32D (30 mg)was dissolved in MeOH (1 ml), then 10% NaOH aqueous solution (0.5 ml)were added under N2, and the reaction mixture was heated to reflux for 1h. The pH of the reaction solution was adjusted to 3 to 4 by adding 1NHCl solution, and 5 ml EA was added for extraction. The organic phasewas concentrated and purified on a column (PE/EA/AcOH=1/1/0.01 elution)to give the title compound 32 (21 mg, yield: 71.7%).

1H NMR (400 MHz, DMSO-d6) δ 8.59 (d, J=2.3 Hz, 1H), 8.31 (dd, J=8.6, 2.4Hz, 1H), 7.86 (d, J=9.1 Hz, 1H), 7.62 (d, J=5.3 Hz, 1H), 7.59 (s, 1H),7.56 (dd, J=5.7, 3.3 Hz, 1H), 7.51 (dd, J=9.0, 7.1 Hz, 1H), 7.46-7.40(m, 2H), 7.25 (d, J=8.6 Hz, 1H), 7.06 (dd, J=5.7, 4.3 Hz, 1H), 5.02 (s,2H), 1.26-1.09 (m, 5H). LCMS (ESI): calculated for C29H19Cl2FN2O5;[M+H]+: 565.1, found: 565.1.

Example 33 Preparation of Sodium6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl) methoxy)naphthalen-2-yl)oxy)nicotinate

An aq. solution of NaOH (30%, 1.44 g, 1.2 eq) was added to a solution ofCompound 1 (4.99 g, 9.12 mmol) in EtOH at r.t. Alter the reactionmixture was heated at reflux for 6 h, it was cooled to r.t. The solidwas collected by filtration, washed with EtOH (10 ml), and dried to givea gray solid (4.07 g, yield: 78.3%).

Example 34 Preparation of Calcium6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinate

To a solution of Compound 35 (1.00 g, 1.76 mmol) in water (10 ml) wasadded a solution of CaCl2 (1.0 g, 20%) in water. White precipitatesformed. After the reaction mixture was stirred at r.t. for 4 h, thesolid was collected by filtration, washed with water (2.0 ml) to givethe product as a white solid (0.80 g, 76.7%).

Example 35 Preparation of2-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)nicotinic acid

The title compound 35 was prepared according to Route D, following theprocedure of Example 22. 1H NMR (400 MHz, DMSO-d6) δ 13.31 (s, 1H),8.29-8.24 (m, 1H), 8.15-8.10 (m, 1H), 7.67 (d, J=9.1 Hz, 1H), 7.64-7.56(m, 3H), 7.53-7.47 (m, 1H), 7.47-7.42 (m, 1H), 7.36 (d, J=2.6 Hz, 1H),7.22-7.17 (m, 1H), 7.06 (d, J=7.4 Hz, 1H), 6.92-6.87 (m, 1H), 4.98 (s,2H), 1.24-1.08 (m, 5H). LCMS (ESI): calculated for C29H20Cl2N2O5;[M+H]+: 547.1, found: 547.1.

Example 36 Preparation of6-((5-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)nicotinic acid

The title compound 36 was prepared according to Route B, following theprocedure of Example 32. 1H NMR (400 MHz, DMSO-d6) δ 13.19 (s, 1H), 8.65(d, J=2.4 Hz, 1H), 8.29 (dd, J=8.6, 2.4 Hz, 1H), 7.77 (d, J=9.1 Hz, 1H),7.65-7.54 (m, 3H), 7.51-7.44 (m, 1H), 7.44-7.34 (m, 2H), 7.19 (dd,J=9.2, 2.4 Hz, 11H), 7.14 (d, J=8.7 Hz, 11H), 7.01 (d, J=7.4 Hz, 1H),5.09 (s, 2H), 1.31-1.07 (m, 6H). LCMS (ESI): calculated forC29H20Cl2N2O5; [M+H]+: 547.1, found: 547.1.

Example 37 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-5-fluoronaphthalen-2-yl)oxy)-2-methylnicotinicacid

The title compound 37 was prepared according to Route B, following theprocedure of Example 32. 1H NMR (400 MHz, DMSO-d6) δ 13.01 (s, 1H), 8.24(d, J=8.6 Hz, 1H), 7.91 (d, J=9.1 Hz, 1H), 7.68 (s, 1H), 7.62 (d, J=9.1Hz, 1H), 7.58 (d, J=7.9 Hz, 2H), 7.55-7.51 (m, 1H), 7.43-7.34 (m, 2H),6.92 (d, J=8.6 Hz, 11H), 5.09 (s, 2H), 2.52 (s, 3H), 1.19-1.08 (m, 4H).LCMS (ESI): calculated for C30H21Cl2FN2O5; [M+H]+: 579.1, found: 579.1.

Example 38 Preparation of6-((7-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-8-fluoronaphthalen-2-yl)oxy)nicotinicacid

The title compound 38 was prepared according to Route B, following theprocedure of Example 32. 1H NMR (400 MHz, DMSO-d6) δ 8.64 (s, 1H), 8.30(dd, J=8.7, 2.4 Hz, 1H), 7.96 (d, J=8.9 Hz, 1H), 7.70 (d, J=9.0 Hz, 1H),7.64-7.48 (m, 3H), 7.37 (t, J=8.8 Hz, 1H), 7.27 (d, J=9.2 Hz, 1H), 7.19(d, J=8.6 Hz, 1H), 6.85 (s, 1H), 5.10 (s, 2H), 2.07-1.89 (m, 1H),0.94-0.76 (m, 4H). LCMS (ESI): calculated for C29H19Cl2FN2O5; [M+H]+:565.1, found: 565.1.

Example 39 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-5-fluoronaphthalen-1-yl)oxy)-2-methylnicotinicacid

The title compound 39 was prepared according to Route B, following theprocedure of Example 32. 1H NMR (400 MHz, CDCl3) δ 7.94 (dd, J=7.9, 5.0Hz, 11H), 7.83 (d, J=8.1 Hz, 1H), 7.72-7.59 (m, 2H), 7.58-7.49 (m, 2H),7.49-7.38 (m, 2H), 7.25-6.97 (m, 1H), 6.67 (d, J=7.9 Hz, 1H), 5.52 (d,J=16.9 Hz, 1H), 5.24 (d, J=16.9 Hz, 1H), 2.70-2.96 (M, 1H), 2.61 (s,3H), 1.05-0.89 (m, 4H). LCMS (ESI): calculated for C30H21Cl2FN2O5;[M+H]+: 579.1, found: 579.1.

Example 40 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)-5-methylpicolinicacid

The title compound 40 was prepared according to Route D, following theprocedure of Example 22. 1H NMR (400 MHz, CDCl3) δ 8.34 (d, J=8.5 Hz,1H), 7.79 (d, J=8.2 Hz, 1H), 7.65-7.55 (m, 2H), 7.54-7.49 (m, 2H),7.50-7.40 (m, 2H), 7.22 (t, J=2.3 Hz, 1H), 7.03-6.93 (m, 2H), 5.44 (s,2H), 2.95-2.58 (m, 1H), 2.22 (s, 3H), 1.01 (m, 4H). LCMS (ESI):calculated for C30H22Cl2N2O5; [M+H]+: 561.1, found: 561.1.

Example 41 Preparation of6-((2,4-dichloro-6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)nicotinic acid

The title compound 41 was prepared according to Route C, following theprocedure of Example 20. 1H NMR (400 MHz, DMSO-d6) δ 13.17 (s, 1H), 8.66(d, J=2.4 Hz, 1H), 8.34 (dd, J=8.6, 2.4 Hz, 1H), 7.84-7.78 (m, 2H),7.70-7.65 (m, 2H), 7.64-7.56 (m, 2H), 7.38 (dd, J=9.1, 2.3 Hz, 1H), 7.24(d, J=8.6 Hz, 1H), 4.92 (s, 2H), 2.45-2.41 (m, 1H), 1.22-1.10 (m, 4H).LCMS (ESI): calculated for C29H18C14N2O5; [M+1]+: 615.0, found: 615.0.

Example 42 Preparation of6-((2-chloro-6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)nicotinic acid

The title compound 42 was prepared according to Route C, following theprocedure of Example 20. 1H NMR (400 MHz, DMSO-d6) δ 13.15 (s, 1H), 8.65(d, J=2.4 Hz, 1H), 8.31 (dd, J=8.6, 2.4 Hz, 1H), 7.76 (d, J=2.4 Hz, 1H),7.70 (d, J=8.8 Hz, 3H), 7.66-7.59 (m, 1H), 7.51 (d, J=8.8 Hz, 1H), 7.44(d, J=9.1 Hz, 1H), 7.22 (dd, J=9.1, 2.4 Hz, 1H), 7.19 (d, J=8.6 Hz, 1H),4.87 (s, 2H), 2.46-2.40 (m, 1H), 1.30-1.09 (m, 4H). LCMS (ESI):calculated for C29H19Cl3N2O5; [M+H]+: 581.0, found: 581.0.

Example 43 Preparation of6-((1-chloro-6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-2-yl)oxy)-2-methylnicotinicacid

The title compound 43 was prepared according to Route C, following theprocedure of Example 20. 1H NMR (400 MHz, DMSO-d6) δ 12.97 (s, 1H), 8.24(d, J=8.6 Hz, 1H), 8.00 (d, J=9.2 Hz, 1H), 7.80 (d, J=8.9 Hz, 1H), 7.60(d, J=8.0 Hz, 2H), 7.55-7.49 (m, 1H), 7.47 (s, 1H), 7.43 (d, J=8.8 Hz,1H), 7.12 (dd, J=9.2, 2.5 Hz, 1H), 6.95 (d, J=8.6 Hz, 1H), 5.02 (s, 2H),2.47 (s, 3H), 1.23-1.11 (m, 5H). LCMS (ESI): calculated forC30H21Cl3N2O5; [M+H]+: 595.1, found: 595.1.

Example 44 Preparation of2-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-2-fluoronaphthalen-1-yl)oxy)-4-fluorobenzoicacid

The title compound 44 was prepared according to Route C, following theprocedure of Example 20. 1H NMR (400 MHz, DMSO-d6) δ 13.04 (s, 1H), 7.82(t, J=8.6 Hz, 1H), 7.71 (d, J=9.2 Hz, 1H), 7.57 (d, J=7.8 Hz, 2H),7.52-7.46 (m, 1H), 7.39 (d, J=2.4 Hz, 1H), 7.37-7.31 (m, 1H), 7.15-7.10(m, 1H), 7.01 (dd, J=9.3, 2.4 Hz, 1H), 6.88 (dd, J=12.3, 2.4 Hz, 1H),6.73 (dd, J=8.7, 2.3 Hz, 1H), 5.08 (s, 211), 1.28-1.05 (m, 4H). LCMS(ESI): calculated for C30H19Cl2F2NO5; [M+H]+: 582.1, found: 582.1.

Example 45 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-2-fluoronaphthalen-1-yl)oxy)-2-methylnicotinicacid

The title compound 45 was prepared according to Route C, following theprocedure of Example 20. 1H NMR (400 MHz, DMSO-d6) δ 12.98 (s, 1H), 8.21(d, J=8.6 Hz, 1H), 7.65 (dd, J=9.3, 1.7 Hz, 1H), 7.57 (d, J=7.7 Hz, 2H),7.51-7.44 (m, 1H), 7.37 (d, J=2.5 Hz, 1H), 7.35-7.29 (m, 1H), 7.16-7.11(m, 1H), 6.98 (dd, J=9.2, 2.5 Hz, 1H), 6.87 (d, J=8.5 Hz, 1H), 5.08 (s,2H), 1.22-1.09 (m, 4H). LCMS (ESI): calculated for C30H21Cl2FN2O5;[M+H]+: 579.1, found: 579.1.

Example 46 Preparation of6-((2-chloro-6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)naphthalen-1-yl)oxy)-2-methylnicotinic acid

The title compound 46 was prepared according to Route C, following theprocedure of Example 20. 1H NMR (400 MHz, DMSO-d6) δ 13.03 (s, 1H), 8.25(d, J=8.6 Hz, 1H), 7.76 (d, J=9.2 Hz, 1H), 7.69 (d, J=8.2 Hz, 1H), 7.58(d, J=8.0 Hz, 2H), 7.50 (d, J=8.1 Hz, 1H), 7.49-7.44 (m, 1H), 7.18 (d,J=8.2 Hz, 1H), 7.04 (dd, J=9.2, 2.4 Hz, 1H), 6.95 (d, J=8.6 Hz, 1H),5.13 (s, 211), 2.59-2.54 (m, 1H), 2.48 (s, 311), 1.27-1.15 (m, 411).LCMS (ESI): calculated for C30H21Cl3N2O5; [M+H]+: 595.1, found: 595.1.

Example 47 Preparation of6-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-2-fluoronaphthalen-1-yl)oxy)-5-methylnicotinicacid

The title compound 47 was prepared according to Route C, following theprocedure of Example 20. 1H NMR (400 MHz, DMSO-d6) δ 13.09 (s, 1H), 8.32(s, 1H), 8.21 (s, 1H), 7.67-7.55 (m, 3H), 7.55-7.45 (m, 1H), 7.39 (s,11H), 7.37-7.30 (m, 1H), 7.20-7.09 (m, 1H), 7.03-6.93 (m, 1H), 5.09 (s,2H), 2.48 (s, 3H), 1.41-1.00 (m, 5H). LCMS (ESI): calculated forC30H21Cl2FN2O5; [M+H]+: 579.1, found: 579.1.

Example 48 Preparation of2-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-2-fluoronaphthalen-1-yl)oxy)nicotinicacid

The title compound 48 was prepared according to Route C, following theprocedure of Example 20. 1H NMR (400 MHz, DMSO-d6) δ 8.29-8.27 (m, 1H),8.22 (d, J=8.5 Hz, 1H), 8.09-8.06 (m, 1H), 7.53-7.47 (m, 3H), 7.46-7.42(m, 1H), 7.30-7.25 (m, 2H), 7.08-7.05 (m, 1H), 6.96 (dd, J=8.4, 2.4 Hz,1H), 5.44 (s, 2H), 2.79 (p, J=6.4 Hz, 1H), 1.20-1.09 (m, 5H). LCMS(ESI): calculated for C29H19Cl2FN2O5; [M+H]+: 565.1, found: 565.1.

Example 49 Preparation of2-((6-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)-2-fluoronaphthalen-1-yl)oxy)-4-fluorobenzoicacid

The title compound 49 was prepared according to Route D, following theprocedure of Example 22. 1 H NMR (400 MHz, DMSO-d6) δ 13.62 (s, 1H),7.84 (d, J=9.2 Hz, 1H), 7.63-7.55 (m, 3H), 7.54-7.49 (m, 1H), 7.42 (t,J=7.9 Hz, 1H), 7.40-7.32 (m, 2H), 7.06 (t, J=8.8 Hz, 1H), 6.96 (dd,J=9.1, 2.5 Hz, 1H), 6.90 (d, J=7.5 Hz, 1H), 6.56 (d, J=8.4 Hz, 1H), 4.99(s, 2H), 1.23-1.09 (m, 4H). LCMS (ESI): calculated for C30H20Cl2FN2O5;[M+H]+: 564.1, found: 564.1.

BIOLOGY EXAMPLES

Compound of Formula (I) in HBV Cell-Based Assay

PHH (Primary Human Hepatocyte) Assay: Cell Line and Compound Treatment

The PHH cells were seeded into the 48-well plates at the density of1.32×105 cells/well. The PHH cells seeding date was defined as day 0.The PHH cells were infected with HBV (D type) at 1600 GE/cell on day 1.The culture medium containing compounds was refreshed on day 2, 4, 6 andday 8. On day 9, the cells were treated with CCK8 and culturesupernatant and cells were collected.

FXR Compounds were tested at 10000.0, 3000.0, 1000.0, 300.0, 100.0,30.0, and 10.0 nM, and ETV was tested at 0.2000, 0.0667, 0.0222, 0.0074,0.0025, 0.0008, and 0.0003 nM. 1% DMSO was used as non-treatmentcontrol. The supernatants on day 9 were collected, and analyzed for HBVDNA by qPCR, HBV RNA by RT-qPCR, HBsAg and HBeA by ELISA.

HBV DNA, RNA and Antigen Assay

DNA in the culture supernatants was isolated with the QIAamp 96 DNABlood Kit according to the manual and quantified by the qPCR. 80 μl ofthe culture supernatants sample was used to extract DNA. The elutionvolume was 120 μl. The PCR mix (8 μl/well) and the samples (2 μl/well)or plasmid standards (2 μl/well) were added to the 384-well opticalreaction plate and performed using the following program: 95C for 10min, then cycling at 95° C. 15 sec, 60° C. 1 min for 40 cycles.

RNA in the culture supernatants was isolated with the PureLink™ Pro 96Viral RNA Kit according to the manual. 35 μl of the culture supernatantssample was used to extract RNA. The elution volume was 100 μl. HBV RNAwas quantified by RT-qPCR.

For HepG2-NTCP Assay, HBsAg and HBeAg in the samples harvested on day 9were measured by the HBsAg ELISA kit (Autobio) and HBeAg ELISA kit(Autobio) according to the manual. For PHH assay, ChemiluminescenceApparatu was used to measure HBsAg according to the Antu HBsAg ELISA kitmanual.

After collecting culture supernatants at the terminal day for eachplate, CCK-8 working solution (diluted with fresh culture medium atratio of 1:9) was added to the cell plates. The plates were incubated at37° C., 5% CO2 incubator for approximate 30 min. The ODs were measuredby microplate reader (OD450 nm/OD630 nm).

Dose Response Curves

% HBV DNA inhibition=(1−HBV DNA copy number of test sample/avg. HBV DNAcopy number of 1% DMSO control)×100%

% HBV RNA inhibition=(1−HBV RNA copy number of test sample/avg. HBV RNAcopy number of 1% DMSO control)×100%

% HBsAg inhibition=(1−HBsAg quantity of sample/HBsAg quantity of 1% DMSOcontrol)×100%

% HBeAg inhibition=(1−HBeAg quantity of sample/HBeAg quantity of 1% DMSOcontrol)×100%

% Cell viability=value of sample/a value of 1% DMSO×100%.

The EC50 and CC50 values were determined by dose-response curves fittedby GraphPad Prism using “log (agonist) vs. response-variable slope”.

Results

As shown in in FIGS. 1, 2 and 3 , in PHH assay, Compound 1, 22, and 27had dose-dependent inhibition of extracellular HBV DNA HBsAg and HBVRNA. As control, ETV showed significant inhibition of HBV DNA, but nosignificant inhibition of HBV HBsAg and HBV RNA.

Compound of Formula (I) in AAV/HBV Animal Model

AAV/HBV Mouse Model and Compound Treatment

Male C57BL′6 mice of 5-week old (obtained from Shanghai Jihui LaboratoryAnimal Care Co., Ltd) were given rAAV8-1.3HBV (1×1011 v.g.) via tailvein to establish infection on Day −28. After 14 days, 21 days, and onDay −4, blood was drawn from submandibular vein to determine the levelof HBV DNA, and HBsAg. On Day-4, 40 mice were chosen based on HBV DNA,and HBsAg results, and were randomly assigned to 5 groups. After themice were infected with HBV virus for 28 days, treatment was givenorally once per day, for 4 weeks. Group 1 was given vehicle (0.5%CMC-Na, 10 ml/kg), Group 2 was given positive control ETV (0.1 mg/kg).Groups 3, 4, and 5 were given Compound 33 at 10 mg/kg, 30 mg/kg, and 60mg/kg, respectively. Blood was drawn from submandibular vein every weekto determine the concentration of HBV DNA, HBV RNA, and HBsAg. On Day28, blood was taken from the heart.

Blood Sample Analysis

HBV DNA determination by qPCR. DNA in the mice plasma was isolated withthe QIAamp 96 DNA Blood Kit according to the manual and quantified bythe qPCR, and performed using the following program: 95C for 10 min,then cycling at 95 V 15 sec, 60° C. 1 min for 40 cycles.

HBV RNA determination by qPCR. RNA in the mice plasma was isolated withthe PureLink™ Pro 96 Viral RNA/DNA Kit according to the manual. 20 μl ofthe plasma was used to extract RNA

HBsAg determination by ELISA. HBsAg in the samples was measured by theHBsAg ELISA kit (Autobio) according to the manual. Briefly, after theplasma was diluted 1200× in a coated plate, and incubate with the enzymeconjugated (37V, 60 min), and the plate was washed 5 time. Substrate wasadded, and it was kept from light at room temperature for 10 min andintensity was measured by a plated reader.

Data Analysis

Average±Standard deviation was calculated for each group, and analyzedwith Student's t-test.

Results

As shown in FIG. 4 , after Compound 33 was given to the mice for 28days, the HBV DNA in the high dose group (60 mg/kg) was significantlylower than that of the vehicle group. On average, the log 10[DNAcopy/uL] of the high dose group was 0.77-1.12 lower than the vehiclegroup (p<0.05).

As shown in FIG. 5 , after Compound 33 was given to the mice for 28days, the HBV RNA in the high dose group (60 mg/kg) was significantlylower than that of the vehicle group. On average, the log 10[RNAcopy/uL] of the high dose group was 0.60-0.66 lower than the vehiclegroup (p<0.01). ETV treatment showed no significant effect on HBV RNA ascompared to the vehicle group.

As shown in FIG. 6 , after Compound 33 was given to the mice for 28days, the HBsAg in the high dose group (60 mg/kg) was lowered by 0.38log 10 [IU/ml] as compared to the vehicle group. ETV treatment showed nosignificant effect on HBsAg as compared to the vehicle group.

1. A method of treating or preventing infection with hepatitis B virusin a human or animal, comprising administering to the human or animal inneed thereof a therapeutically effective amount of a phenylisoxazolylmethylene-naphthalene-ether derivative having a structure of formula(I), or a pharmaceutically acceptable salt, ester or stereoisomerthereof:

wherein: R¹, R² and R³ are independently selected from H, halogen,unsubstituted or halogen substituted C₁₋₆ alkyl and unsubstituted orhalogen substituted C₁₋₆ alkoxy, provided that at least one of R¹, R²and R³ is not hydrogen, R⁰ is selected from unsubstituted or halogensubstituted C₁₋₆ alkyl, C₃₋₆ cycloalkyl and C₄₋₇ alkylcycloalkyl; X¹ andX² are independently selected from H and halogen; moiety —O—Z attachesto the naphthalene ring, wherein Z is a residue selected from 5-10membered aryl or 5-10 membered heteroaryl optionally having one or morehetero atoms selected from N, O and S, wherein the 5-10 membered aryl or5-10 membered heteroaryl is substituted by R⁴ and is optionally furthersubstituted by R⁵; and wherein R4 is selected from —COOH, —CH₂COOH,—NHSO₂CF₃, —SO₂NH—C₁₋₆ alkyl, —SO₃H, —CONHSO₂—C₁₋₆ alkyl, —CONHSO₂—C₃₋₆cycloalkyl, —CONHSO₂-5-10 membered aryl and —CONHSO2-5-10 membered arylsubstituted by C₁₋₆ alkyl at the aryl, and wherein R⁵ is selected fromH, C₁₋₆ alkyl, halogen, C₁₋₆ haloalkyl, —O—(C₁₋₆ alkyl) and —NH—(C₁₋₆alkyl).
 2. The method according to claim 1, wherein R¹, R² and R³ areindependently selected from H, halogen and C₁₋₃ perfluoroalkoxy, and R⁰is selected from isopropyl or cyclopropyl; wherein Z is a phenyl whichis substituted by R⁴ and is optionally substituted by R⁵; or Z is a 5-10membered heteroaryl having one or more hetero atoms selected from N, Oand S, which is substituted by R⁴ and is optionally substituted by R⁵;and wherein the halogen is fluoro or chloro.
 3. The method according toclaim 2, wherein R¹, R² and R³ are independently selected from H, Cl, Fand —O—CF₃; and wherein Z is a 5-6 membered heteroaryl having one ormore hetero atoms selected from N, O and S, which is substituted by R⁴and is optionally substituted by R⁵, wherein R⁴ is selected from —COOH,—CH₂COOH, —CONHSO₂—C₁₋₆ alkyl and —CONHSO₂—C₃₋₆ cycloalkyl, R⁵ isselected from H, C₁₋₃ alkyl and halogen.
 4. The method according toclaim 3, wherein Z is pyridyl; R⁴ is —COOH; and R⁵ is H or halogen. 5.The method according to claim 1, wherein the phenylisoxazolylmethylene-naphthalene-ether derivative is of one of the followingstructures:


49. The method according to claim 4, wherein the phenylisoxazolylmethylene-naphthalene-ether derivative is of the following structure:


7. The method according to claim 1, wherein the method comprisesadministering the phenylisoxazolylmethylene-naphthalene-ether derivativehaving the structure of formula (I), or a pharmaceutically acceptablesalt, ester or stereoisomer thereof in combination with one or moreother anti-HBV agents.
 8. The method of claim 7, wherein the otheranti-HBV agents are selected from HBV vaccines, HBV DNA polymeraseinhibitors, immunomodulators, toll-like receptor (TLR) modulators,interferon alpha receptor ligands, hepatitis b surface antigen (HBsAg)inhibitors, cyclophilin inhibitors, HBV viral entry inhibitors,antisense oligonucleotide targeting viral mRNA, short interfering RNAs(siRNA) and ddRNAi endonuclease modulators, ribonucleotide reductaseinhibitors, HBV E antigen inhibitors, covalently closed circular DNA(cccDNA) inhibitors, fatty acid synthase inhibitors, HBV antibodies,CCR2 chemokine antagonists, retinoic acid-inducible gene 1 stimulators,NOD2 stimulators, PD-1 inhibitors, PD-L1 inhibitors, KDM inhibitors, andHBV replication inhibitors.
 9. The method of claim 8, wherein the HBVDNA polymerase inhibitor is entecavir or tenofovir, the PD-1 inhibitoris one or more selected from nivolumab, pembrolizumab, pidilizumab,BGB-108, SHR-1210, PDR-001, PF-06801591, IBI-308, cemiplimab,camrelizumab, sintilimab, tislelizumab (BGB-A317), BCD-100,JNJ-63723283, Zimberelimab (GLS-010, WBP-3055), Balstilimab (AGEN2034)and dostarlimab (TSR-042); the PD-L1 inhibitors is one or more selectedfrom atezolizumab (RG-7446), avelumab, BGB-A333, BMS-936559 (MDX-1105),durvalumab, CX-072, GX-P2, KN035 (ASC022), GS-4224 and INCB086550; theantisense oligonucleotide is Ionis-HBVRx or Ionis-HBV-LRx the shortinterfering RNA is JNJ-3989, or Vir-2218, or DCR-HBVS the fatty acidsynthase inhibitor is TVB-2640 and/or TVB-3567; the interferon isPegasys; and the capsid inhibitor is one or more selected fromABI-H0731, ABI-H2158, ABI-H3733, CB-HBV-001, JNJ-6379, JNJ-0440, QL-007,RG-7907 and RO7049389.
 10. (canceled)
 11. (canceled)
 12. (canceled) 13.(canceled)
 14. (canceled)
 15. (canceled)
 16. A pharmaceuticalcomposition for anti-hepatitis B virus, comprising a therapeuticallyeffective amount of a phenylisoxazolyl methylene-naphthalene-etherderivative having a structure of formula (I), or a pharmaceuticallyacceptable salt, ester or stereoisomer thereof and one or more otheranti-HBV agents, and a pharmaceutically acceptable auxiliary material,

wherein: R¹, R² and R³ are independently selected from H, halogen,unsubstituted or halogen substituted C₁₋₆ alkyl and unsubstituted orhalogen substituted C₁₋₆ alkoxy, provided that at least one of R¹, R²and R³ is not hydrogen, R⁰ is selected from unsubstituted or halogensubstituted C₁₋₆ alkyl, C₃₋₆ cycloalkyl and C₄₋₇ alkylcycloalkyl; X¹ andX2 are independently selected from H and halogen; moiety —O—Z attachesto the naphthalene ring, wherein Z is a residue selected from 5-10membered aryl or 5-10 membered heteroaryl optionally having one or morehetero atoms selected from N, O and S, wherein the 5-10 membered aryl or5-10 membered heteroaryl is substituted by R⁴ and is optionally furthersubstituted by R⁵; and wherein R⁴ is selected from —COOH, —CH₂COOH,—NHSO₂CF₃, —SO₂NH—C₁₋₆ alkyl, —SO₃H, —CONHSO₂—C₁₋₆ alkyl, —CONHSO₂—C₃₋₆cycloalkyl, —CONHSO₂-5-10 membered aryl and —CONHSO₂-5-10 membered arylsubstituted by C₁₋₆ alkyl at the aryl, and wherein R⁵ is selected fromH, C₁₋₆ alkyl, halogen, C₁₋₆ haloalkyl, —O—(C₁₋₆ alkyl) and —NH—C₁₋₄alkyl).
 17. The pharmaceutical composition according to claim 16,wherein R¹, R² and R³ are independently selected from H, halogen andC₁₋₃ perfluoroalkoxy, and R⁰ is selected from isopropyl or cyclopropyl;wherein Z is a phenyl which is substituted by R⁴ and is optionallysubstituted by R⁵; or Z is a 5-10 membered heteroaryl having one or morehetero atoms selected from N, O and S, which is substituted by R⁴ and isoptionally substituted by R⁵; and wherein the halogen is fluoro orchloro.
 18. The pharmaceutical composition according to claim 17,wherein R¹, R² and R³ are independently selected from H, Cl, F and—O—CF₃; and wherein Z is a 5-6 membered heteroaryl having one or morehetero atoms selected from N, O and S, which is substituted by R⁴ and isoptionally substituted by R⁵, wherein R is selected from —COOH,—CH₂COOH, —CONHSO₂—C₁₋₆ alkyl and —CONHSO₂—C₃₋₆ cycloalkyl, R⁵ isselected from H, C₁₋₃ alkyl and halogen.
 19. The pharmaceuticalcomposition according to claim 18, wherein Z is pyridyl; R⁴ is —COOH;and R⁵ is H or halogen.
 20. The pharmaceutical composition according toclaim 16, wherein the phenylisoxazolyl methylene-naphthalene-etherderivative is of one of the following structures:


21. The pharmaceutical composition according to claim 16, wherein thephenylisoxazolyl methylene-naphthalene-ether derivative is of thefollowing structure:


22. The pharmaceutical composition according to claim 16, wherein thepharmaceutical composition comprises one or more other anti-HBV agentsand the phenylisoxazolylmethylene-naphthalene-ether derivative havingthe structure of formula (I), or a pharmaceutically acceptable salt,ester or stereoisomer thereof.
 23. The pharmaceutical composition ofclaim 22, wherein the other anti-HBV agents are selected from HBVvaccines, HBV DNA polymerase inhibitors, immunomodulators, toll-likereceptor (TLR) modulators, interferon alpha receptor ligands, hepatitisb surface antigen (HBsAg) inhibitors, cyclophilin inhibitors, HBV viralentry inhibitors, antisense oligonucleotide targeting viral mRNA, shortinterfering RNAs (siRNA) and ddRNAi endonuclease modulators,ribonucleotide reductase inhibitors, HBV E antigen inhibitors,covalently closed circular DNA (cccDNA) inhibitors, fatty acid synthesisinhibitors, HBV antibodies, CCR2 chemokine antagonists, retinoicacid-inducible gene 1 stimulators, NOD2 stimulators, PD-1 inhibitors,PD-L1 inhibitors, KDM inhibitors, and HBV replication inhibitors. 24.The pharmaceutical composition of claim 23, wherein the HBV DNApolymerase inhibitor is entecavir or tenofovir, the PD-1 inhibitor isone or more selected from nivolumab, pembrolizumab, pidilizumab,BGB-108, SHR-1210, PDR-001, PF-06801591, IBI-308, cemiplimab,camrelizumab, sintilimab, tislelizumab (BGB-A317), BCD-100,JNJ-63723283, Zimberelimab (GLS-010, WBP-3055), Balstilimab (AGEN2034)and dostarlimab (TSR-042); the PD-L1 inhibitors is one or more selectedfrom atezolizumab (RG-7446), avelumab, BGB-A333, BMS-936559 (MDX-1105),durvalumab, CX-072, GX-P2, KN035 (ASC022), GS-4224 and INCB086550; theantisense oligonucleotide is Ionis-HBVRx or Ionis-HBV-LRx; the shortinterfering RNA is JNJ-3989, Vir-2218, or DCR-HBVS; the fatty acidsynthase inhibitor is TVB-2640 and/or TVB-3567; the interferon isPegasys; and the capsid inhibitor is one or more selected fromABI-H0731, ABI-H2158, ABI-H3733, CB-HBV-001, JNJ-6379, JNJ-0440, QL-007,RG-7907 and RO7049389.